3-aryl-3-hydroxy-2-amino-propionic acid amides, 3-heteroaryl-3-hydroxy-2-amino-propionic acid amides and related compounds having analgesic and/or immuno stimulant activity

ABSTRACT

Compounds of Formulas 1 and 2 
     
       
         
         
             
             
         
       
     
     where the variables have the meaning disclosed in the specification, have analgesic and in some cases immunostimulant activity.

CLAIM OF PRIORITY

This is a national stage application under 35 U.S.C. §371 of PCTapplication PCT/US2006/002570, filed on Jan. 25, 2006, which claims thebenefit of Provisional Application No. 60/647,271, filed on Jan. 26,2005.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to derivatives of3-aryl-3-hydroxy-2-amino-propionic acid amides,3-heteroaryl-3-hydroxy-2-amino-propionic acid amides and to relatedcompounds having analgesic and in some cases immuno stimulant activity.

The present invention also relates to pharmaceutical compositionscontaining these compounds as active ingredient for alleviating oreliminating pain in mammals and/or stimulating the immune system inmammals and to methods of using said pharmaceutical compositions asanalgesics and or immuno stimulants.

2. Background Art

Several compounds falling within one or more of the general definitionsas “derivatives of 3-aryl-3-hydroxy-2-amino-propionic acid amides, of3-heteroaryl-3-hydroxy-2-amino-propionic acid amides, of1-aryl-1-hydroxy-2,3-diamino-propyl amines,1-heteroaryl-1-hydroxy-2,3-diamino-propyl amines” are known in thepatent and scientific literature.

For example, United States Patent Application Publications US2003/0153768; US 2003/0050299 disclose several examples of theabove-mentioned known compounds. The N-acyl compounds of thesereferences are said to be useful as N-acylsphingosineglucosyltransferase inhibitors, the amide and the reduced compounds aredescribed as intermediates in their preparations. Illustrative specificexamples of compounds of these references are shown below:

The publication Shin et al. Tetrahedron Asymmetry, 2000, 11, 3293-3301discloses the following compounds:

L-threo-PDMP and some other known compounds used in the methods of thisinvention are commercially available, in pure enantiomeric and racemicforms, as applicable, from Matreya, LLC Pleasant Gap, Pa.

U.S. Pat. Nos. 5,945,442; 5,952,370; 6,030,995 and 6,051,598, which areall related to each other as being based on same or related disclosures,describe compounds which are structurally similar to the known compoundsshown above. The compounds of these U.S. patent references are said tobe inhibitors of the enzyme glucosylceramide (GlcCer) synthethase.

A publication in Journal of Labelled Compounds & Radiopharmaceuticals(1996), 38(3), 285-97 discloses the compound of the formula

Published PCT application WO 01/38228 discloses

in connection with a chromatographic method.

Kastron et al. in Latvijas PSR Zinatnu Akademijas Vestis, Kimijas Serija(1965) (4), 474-7 disclose the following compound.

Significantly, according to the best knowledge of the present inventors,none of the compounds of the prior art which are structurally similar tothe novel compounds of the present invention are known in the prior artas analgesics or immunostimulants.

SUMMARY OF THE INVENTION

The present invention is directed to compounds of Formula 1

where R₁ is H or alkyl of 1 to 6 carbons,R₂ is H, alkyl of 1 to 6 carbons or the R₁ and R₂ groups together withthe nitrogen form a saturated or unsaturated 4, 5, 6 or 7 membered ringthat optionally includes one or two heteroatoms independently selectedfrom N, O and S, said 4, 5, 6 or 7 membered ring optionally beingsubstituted with one or two COOH, CH₂OH, OH, B(OH)₂, cyano or halogengroups or with one or two alkyl groups having 1 to 6 carbons, or one ortwo carbons of said rings being attached to an oxygen to form ketogroups and said 4, 5, 6 or 7 membered ring optionally being condensedwith an aromatic or non-aromatic 5 or 6 membered ring that optionallyincludes 1 or heteroatoms selected from N, O and S;R₃ is independently selected from H, alkyl of 1 to 20 carbons,cycloalkyl of 3 to 6 carbons, aryl or heteroaryl, aryl-alkyl,aryl-(hydroxy)alkyl, heteroaryl-alkyl or hetero-(hydroxy)alkyl where thealkyl moiety has 1 to 4 carbons, said aryl or heteroaryl groups beingoptionally substituted with 1 to 3 groups independently selected fromthe group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to6 carbons and thioxy of 1 to 6 carbons, or R₃ is CO—R₇, SO₂R₇ or CO—O—R₇where R₇ is H, alkyl of 1 to 1 to 20 carbons, alkyl of 1 to 20 carbonssubstituted with and NH₂ group or with an NH—COalkyl group where thealkyl group has one to 6 carbons, aryl or heteroaryl, aryl-alkyl orheteroaryl-alkyl where the alkyl moiety has 1 to 4 carbons, said aryl orheteroaryl groups being optionally substituted with 1 to 3 groupsindependently selected from the group consisting of halogen, alkyl of 1to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 6 carbons;R₄ is H, alkyl of 1 to 6 carbons or CO—R₈ where R₈ is alkyl of 1 to 6carbons; the wavy lines represent bonds connected to carbons having R orS configuration;the dashed lines represent a bond or absence of a bond with the provisothat the ring containing the dashed lines is aromatic;m, n and q are integers independently selected from 0, 1, 2 or 3 withthe proviso that the sum of m, n and q is 2 or 3;s is zero (0) or when X is N then s is zero (0) or 1; W, X and Yindependently represent a CH, CR₅, CR₆ or a heteroatom selectedindependently of N, O and S, andR₅ and R₆ are independently selected from H, halogen, alkyl of 1 to 6carbons, halogen substituted alkyl of 1 to 6 carbons, alkoxy of 1 to 6carbons and thioxy of 1 to 6 carbons, phenyl, orR₅ and R₆ together with the atoms to which they are attached jointlyform a carbocyclic or a heterocyclic ring, the carbocyclic ring having 5or 6 atoms in the ring, the heterocyclic ring having 5 or 6 atoms in thering and 1 to 3 heteroatoms independently selected from N, O and S;said carbocyclic or heterocyclic ring jointly formed by R₅ and R₆ beingoptionally substituted with 1 to 6 R₉ groups where R₉ is independentlyselected from halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbonsand thioxy of 1 to 6 carbons or a pharmaceutically acceptable salt ofsaid compoundwith the proviso that Formula 1 does not cover compounds where R₄ is H,R₁ and R₂ jointly with the nitrogen form a pyrrolidino or morpholinoring, the sum of m, n and q is 3, and none of W, X and Y represent aheteroatom with the further proviso that the formula does not cover thecompounds of the formula below:

The present invention is also directed to the compounds of Formula 2

where R₁ is H or alkyl of 1 to 6 carbons,R₂ is H, alkyl of 1 to 6 carbons or the R₁ and R₂ groups together withthe nitrogen form a saturated or unsaturated 4, 5, 6 or 7 membered ringthat optionally includes one or two heteroatoms independently selectedfrom N, O and S, said 4, 5, 6 or 7 membered ring optionally beingsubstituted with one or two COOH, CH₂OH, OH, B(OH)₂, cyano or halogengroups or with one or two alkyl groups having 1 to 6 carbons, or one ortwo carbons of said rings being attached to an oxygen to form ketogroups and said 4, 5, 6 or 7 membered ring optionally being condensedwith an aromatic or non-aromatic 5 or 6 membered ring that optionallyincludes 1 or heteroatoms selected from N, O and S;R₃ is independently selected from H, alkyl of 1 to 20 carbons,cycloalkyl of 3 to 6 carbons, aryl or heteroaryl, aryl-alkyl,aryl-(hydroxy)alkyl, heteroaryl-alkyl or hetero-(hydroxy)alkyl where thealkyl moiety has 1 to 4 carbons, said aryl or heteroaryl groups beingoptionally substituted with 1 to 3 groups independently selected fromthe group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to6 carbons and thioxy of 1 to 6 carbons, or R₃ is CO—R₇, SO₂R₇ or CO—O—R₇where R₇ is H, alkyl of 1 to 1 to 20 carbons, alkyl of 1 to 20 carbonssubstituted with an NH₂, NHCOR₇ or NHCOOR₇ group, aryl or heteroaryl,aryl-alkyl or heteroaryl-alkyl where the alkyl moiety has 1 to 4carbons, said aryl or heteroaryl groups being optionally substitutedwith 1 to 3 groups independently selected from the group consisting ofhalogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of1 to 6 carbons;the wavy lines represent bonds connected to carbons having R or Sconfiguration;the dashed lines represent a bond or absence of a bond with the provisothat the ring containing the dashed lines is aromatic;R₉ and R₁₀ are independently H, alkyl of 1 to 6 carbons or OR₁₁, or R₉and R₁₀ jointly represent NOR₁₁ with the proviso that when the dashedlines between carbons 2 and 3 of the propionic acid moiety represents abond then R₁₀ does not exist and R₉ is not OR₁₁ with the further provisothat when R₉ is OR₁₁ then R₁₀ is not hydrogen;R₁₁ is H, alkyl of 1 to 6 carbons or CO—R₁₂ where R₁₂ is alkyl of 1 to 6carbons;m, n and q are integers independently selected from 0, 1, 2 or 3 withthe proviso that the sum of m, n and q is 2 or 3;s is zero (0) or when X is N then s is zero (0) or 1;W, X and Y independently represent a CH, CR₅, CR₆ or a heteroatomselected independently of N, O and S, andR₅ and R₆ are independently selected from H, halogen, alkyl of 1 to 6carbons, halogen substituted alkyl of 1 to 6 carbons, alkoxy of 1 to 6carbons and thioxy of 1 to 6 carbons, phenyl, orR₅ and R₆ together with the atoms to which they are attached jointlyform a carbocyclic or a heterocyclic ring, the carbocyclic ring having 5or 6 atoms in the ring, the heterocyclic ring having 5 or 6 atoms in thering and 1 to 3 heteroatoms independently selected from N, O and S;said carbocyclic or heterocyclic ring jointly formed by R₅ and R₆ beingoptionally substituted with 1 to 6 R₉ groups where R₉ is independentlyselected from halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbonsand thioxy of 1 to 6 carbons or a pharmaceutically acceptable salt ofsaid compound.

The present invention is also directed to pharmaceutical compositionscontaining the above-noted novel compound to be used as analgesicsand/or immunostimulants in mammals, and to methods of using saidpharmaceutical compositions as analgesics or immunostimulants.

DETAILED DESCRIPTION OF THE INVENTION

A general description of the compounds of the invention is provided inthe Summary Section of the present application for patent. Mostcompounds of the invention contain one or more asymmetric centers, suchthat the compounds may exist in enantiomeric as well as indiastereomeric forms. In fact, most of the compounds of the presentinvention have two asymmetric carbons adjacent to one another andtherefore can exist in erythro or threo form, with each of these twoforms having dextrorotatory (D) or levorotary (L) enantiomers. Althoughthe threo form is generally preferred in accordance with the presentinvention for analgesic activity, unless it is specifically notedotherwise, the scope of the present invention includes all enantiomers,diastereomers and diastereomeric or racemic mixtures. In light of theforegoing, it should be clearly understood that the designation “DL” or“(+/−)” or “(±)” in this application includes the pure dextrorotatoryenantiomer, the pure levorotatory enantiomer and all racemic mixtures,including mixtures where the two enantiomers are present in equal or inunequal proportions. Moreover, for simplicity sake in many of thestructural formulas, such as in the example below, only one of theenantiomers is actually shown but when the designation “DL” or “(+/−)”or or “(±)” appears it also includes the enantiomeric form (mirrorimage) of the structure actually shown in the formula.

For Example:

Thus, in the example above, only one enantiomer is shown, but becausethe designation “DL” (or or “(+/−)” or “(±)”) appears below the formula,its optical isomer

and all racemic mixtures of the two optical isomers are also included.

In the case of some compounds of the present invention one enantiomer ofthe threo, and in some cases of the erythro, enantiomers issignificantly more active as an analgesic than the other enantiomer ofthe same pair. For this reason the isolated enantiomer which issignificantly more active than the other is considered a novel andinventive composition even if the racemic mixture or the other oppositeenantiomer of the same compound have already been described in the priorart.

Some of the novel compounds of the present invention contain three ormore asymmetric centers. An example is the following compound

named Compound 214 in the description. The formula shown in thedescription for Compound 214 indicates two compounds of the threoisomer, but the two compounds indicated are not mirror images of eachother, they are diastereomers. Another isomer pair is shown anddescribed as Compound 215.

Keeping the foregoing examples in mind the reader one of ordinary skillin the art should readily understand the scope of each describedexample, although in a broad sense all isomers, enantiomers and racemicmixtures are within the scope of the invention.

The term “alkyl” in the general description and definition of thecompounds includes straight chain as well as branch-chained alkylgroups.

Generally speaking the compounds of the invention may form salts withpharmaceutically acceptable acids or bases, and such pharmaceuticallyacceptable salts of the compounds of Formula 1 and of Formula 2 are alsowithin the scope of the invention.

Referring now to the novel compounds of Formula 1, in a class ofpreferred compounds of the invention none of the W, X and Y groups is aheteroatom. Within this class, compounds are preferred where the sum ofm, n and q is 3 and the aromatic group is unsubstituted or substitutedwith one or more halogen, alkyl of 1 to 6 carbons, or halogensubstituted alkyl of 1 to 6 carbons. Compounds within this class arealso preferred where the R₅ and R₆ groups form a carbocyclic ring, or aheterocyclic ring.

In another class of preferred compounds in accordance with Formula 1 oneof the variables W, X and Y represents a heteroatom, preferably nitrogenand the sum of m, n and q is 3.

In still another class of preferred compounds in accordance with Formula1 one or two of the variables W, X and Y represent a heteroatom,selected from N, O or S and the sum of m, n and q is 2.

Referring still to the compounds of Formula 1, compounds are preferredwhere R₄ is H or an acyl group, more preferably H.

With reference to the variables R₃, compounds in accordance with Formula1 are preferred where both R₃ groups are Hand where one R₃ group is Hand the other is benzyl, monohalogeno, dihalogeno, methyl or methoxysubstituted benzyl, cyclohexyl, an alkyl of 1 to 7 carbons, COR₇, COOR₇where R₇ is alkyl of 1 to 15 carbons, benzyloxy, phenyl, methoxyphenyl,monohalogen or dihalogeno substituted phenyl, a 2-hydroxy-1-phenylethylgroup or an alkyl group of 1 to 20 carbons itself substituted with anNH₂, NHCOR₇, or NHCOOR₇ group.

Referring now to the variables R₁ and R₂ in the compounds of Formula 1,compounds are preferred in accordance with the invention where R₁ and R₂jointly form a pyrrolidine, a 3-fluoro or a 3,3-difluoro or an 3-hydroxysubstituted pyrrolidine, a morpholine, a thiomorpholine, a piperazine,an alkyl substituted piperazine where the alkyl group has 1 to 6carbons, an azetidine, a tetrahydrothiazole, an indoline, or a 2H-pyrrolring or R₁ and R₂ are two alkyl groups of 1 to 3 carbons.

Referring now to the novel compounds of Formula 2, with respect to thevariables W, X, Y, m, n, q, R₁, R₂, R₅, R₆, R₃ compounds are gerenerallypreferred in which these variables have the same preferences as incompounds of Formula 1.

With respect to R₉ and R₁₀, compounds are generally preferred where R₉and R₁₀ are both hydrogen, where one of these two variables is hydroxyand the other is alkyl of 1 to 6 carbons, where the R₉ and R₁₀ groupsjointly form an NOR₁₁ group, and where R₉ is hydrogen, the dashed linebetween carbons 2 and 3 represent a double bond and R₁₀ does not exist.With respect to R₁₁ compounds of Formula 2 are preferred where R₁₁ is H,or COR₁₂ where R₁₂ is alkyl of 1 to 3 carbons.

Presently still more preferred are Compounds of Formula 2 where R₁ andR₂ jointly with the nitrogen form a five-membered ring, where both R₃groups are hydrogen and where one of the R₃ groups is hydrogen and theother is formyl.

The presently most preferred novel compounds of the invention aredisclosed with their structural formulas in the ensuing Tables and ordescription, showing activity of exemplary compounds relevant to theirability to act as analgesics and/or immunostimulants.

Biological Activity, Modes of Administration

The novel compounds of the invention have analgesic and/orimmunostimulant activity in mammals. Some of the compounds described inthe introductory section which per se are known in the art, have beendiscovered by the present inventors to also have analgesic effect inmammals. To the best of the knowledge of the present inventors theanalgesic or immunostimulant biological activity of the known compoundswas not known before the present discovery.

An art-accepted model or assay for measuring an analgesic effect of acompound in chronic pain (in particular peripheral neuropathy) is themodel known as Kim and Chung 1992, Pain 150, pp 355-363 (Chung model).This model involves the surgical ligation of the L5 (and optionally theL6) spinal nerves on one side in experimental animals. Rats recoveringfrom the surgery gain weight and display a level of general activitysimilar to that of normal rats. However, these rats developabnormalities of the foot, wherein the hindpaw is moderately everted andthe toes are held together. More importantly, the hindpaw on the sideaffected by the surgery appears to become sensitive to low-thresholdmechanical stimuli and will perceive pain instead of the faint sensationof touch. This sensitivity to normally non-painful touch, called“tactile allodynia”, develops within the first week after surgery andlasts for at least two months. The allodynia response includes liftingthe affected hindpaw to escape from the stimulus, licking the paw andholding it in the air for many seconds. None of these responses isnormally seen in the control group.

To produce the tactile allodynia, rats are anesthetized before surgery.The surgical site is shaved and prepared either with betadine orNovacaine. Incision is made from the thoracic vertebra XIII down towardthe sacrum. Muscle tissue is separated from the spinal vertebra (leftside) at the L4-S2 levels. The L6 vertebra is located and the transverseprocess is carefully removed with a small rongeur to expose the L4-L6spinal nerves. The L5 and L6 spinal nerves are isolated and tightlyligated with 6-0 silk thread. The same procedure is done on the rightside as a control, except no ligation of the spinal nerves is performed.

After a complete hemostasis is confirmed, the wounds are sutured. Asmall amount of antibiotic ointment is applied to the incised area, andthe rat is transferred to the recovery plastic cage under a regulatedheat-temperature lamp.

On the day of the experiment, at least seven days after the surgery,typically six rats per test group are administered the test drugs byintraperitoneal (i.p.) injection or oral gavage (p.o.). For i.p.administration, the compounds are formulated in H₂O and given in avolume of 1 ml/kg body weight by injecting into the intraperitonealcavity. For p.o. administration, the compounds are formulated in H₂O andgiven in a volume of 1 ml/kg body weight using an 18-gauge, 3 inchgavage needle that is slowly inserted through the esophagus into thestomach.

Tactile allodynia is assessed via von Frey hairs, which are a series offine hairs with incremental differences in stiffness. Rats are placed ina plastic cage with a wire mesh bottom and allowed to acclimate forapproximately 30 minutes. To establish the pre-drug baseline, the vonFrey hairs are applied perpendicularly through the mesh to themid-plantar region of the rats' hindpaw with sufficient force to causeslight buckling and held for 6-8 seconds. The applied force has beencalculated to range from 0.41 to 15.1 grams. If the paw is sharplywithdrawn, it is considered a positive response. A normal animal willnot respond to stimuli in this range, but a surgically ligated paw willbe withdrawn in response to a 1-2 gram hair. The 50% paw withdrawalthreshold is determined using the method of Dixon, W. J., Ann. Rev.Pharmacol. Toxicol. 20:441-462 (1980) hereby incorporated by reference.Tactile allodynia is measured prior to and 15, 30, and 60 minutes afterdrug administration. The post-drug threshold is compared to the pre-drugthreshold and the percent reversal of tactile sensitivity is calculatedbased on a normal threshold of 15.1 grams.

Table 1 below indicates the degree of pain reversal obtained in theChung model with exemplary compounds of the invention. Theintraperitonial (i.p.) and/or intravenous (iv) administration of thecompounds was in doses ranging from 1 μg/kg to 300 μg/kg or 3 mg/kg POand the peak percentage of reversal of allodynia was measured at 15, 30or 60 minutes after administration, as is indicated in the table. Dataare expressed as the highest % allodynia reversal (out of 3 time points:15 min, 30 min, or 60 min. post-drug) with a minimum of a 20% allodyniareversal in the rat Chung model. Comparisons between groups (drugtreated vs. saline treated) were made using a two-tailed, 2-sample,unpaired t-test. Compounds that are not shown which were notstatistically analgesic following an IP dose of 300 ug/kg, but may stillbe analgesic. Compounds that do not exhibit significant analgesia at 100mg/kg are not considered to be analgesic.

TABLE 1 Peak % Pain Dose μg/kg, Compound reversal: time Mode of #Chemical Formula post dose administ.  22

100% 60 min 3000 μg/kg PO  22

100% 60 min 100 μg/kgIP  20

92%60 min  30 μg/kgIP  35

92%60 min 300 μg/kgIP  23

100% 60 min  30 μg/kgPO  24

60%60 min 300 μg/kgIP  58

75%60 min 300 μg/kgIP  59

92%60 min 300 μg/kgIP  27

42%30 min 300 μg/kgIP  29

47%60 min 300 μg/kgIP  61

64%60 min 300 μg/kgIP  34

62%60 min 300 μg/kgIP  30

53%60 min 300 μg/kgIP  64

100% 30 min 300 μg/kgIP  55

58%60 min 300 μg/kgIP  56

67%60 min 300 μg/kgIP  67

78%60 min 300 μg/kgIP  68

94%60 min 300 μg/kgPO  69

63%30 min 300 μg/kgIP  41

70%60 min 300 μg/kgIP  49

85%60 min 100 μg/kgIP  43

96%60 min 300 μg/kgIP  26

92%60 min 300 μg/kgIP  57

51%30 min 300 μg/kgIP  28

92%60 min 300 μg/kgIP Compound216

100% 60 min 300 μg/kgIP Compound234

59%60 min 300 μg/kgIP Compound230

52%30 min 300 μg/kgIP Compound236

32%60 min 300 μg/kgIP Compound218

32%60 min 300 μg/kgIP Compound239

75%30 min 300 μg/kgIP Compound238

61%60 min 300 μg/kgIP Compound205

62%30 min 300 μg/kgIP Compound206

67%30 min 300 μg/kgIP Compound240

70%60 min 300 μg/kgIP Compound232

80%60 min 300 μg/kgIP Compound220

32%30 min 300 μg/kgIP Compound210

78%60 min 300 μg/kgIP Compound221

87%60 min 300 μg/kgIP Compound227

95%30 min 300 μg/kgIP Compound226

95%60 min 300 μg/kgIP Compound207

96%60 min 300 μg/kgIP Compound213

85%30 min 300 μg/kgIP Compound214

86%60 min  30 μg/kgIP Compound228

36%60 min 300 μg/kgIP Compound229

53%60 min 300 μg/kgIP Compound224

51%60 min 300 μg/kgIP Compound215

73%60 min 300 μg/kgIP Compound219

82%60 min  30 μg/kgIP Compound203

87%60 min 300 μg/kgIP Compound204

50%60 min 300 μg/kgIP Compound40

47%60 min 300 μg/kgIP Compound247

42%60 min 300 μg/kgIP Compound254

62%60 min 300 μg/kgIP Compound248

59%60 min  30 μg/kgIP Compound255

60%60 min 300 μg/kgIP Compound256

62%60 min 300 μg/kgIP

An art accepted method for measuring immuno stimulation comprisessystemic administration of compounds to assay for the ability tostimulate the immune system, possibly due to nonspecific upregulation ofthe hemolymphoreticular system. This upregulation could result inincreased numbers of lymphocytes of both T- and B-cell lineage. Althoughapplicant does not wish to be bound by the biological theory of theimmunostimulation, actual immunostimulatory efficacy of the compoundscan be demonstrated in vivo by assaying splenic size in response toadministration of the test compound to laboratory test species rats.Animals dosed at 200 mg/kg of Compound 22 of this invention exhibited atwentyfive percent increase in spleen size which demonstratesimmunostimulatory potential of the compound. Generally speaking anycompound that exhibits splenic enlargement following dosing of 200 mg/kgor less may be considered an immunostimulant.

Modes of Administration:

The compounds of the invention may be administered at pharmaceuticallyeffective dosages. Such dosages are normally the minimum dose necessaryto achieve the desired therapeutic effect; in the treatment of chromicpain, this amount would be roughly that necessary to reduce thediscomfort caused by the pain to tolerable levels. For human adults suchdoses generally will be in the range 0.1-5000 mg/day; more preferably inthe range 1 to 3000 mg/day, still more preferably in the range of 10 mgto 1000 mg/day. However, the actual amount of the compound to beadministered in any given case will be determined by a physician takinginto account the relevant circumstances, such as the severity of thepain, the age and weight of the patient, the patient's general physicalcondition, the cause of the pain, and the route of administration.

The compounds are useful in the treatment of pain in a mammal;particularly a human being. Preferably, the patient will be given thecompound orally in any acceptable form, such as a tablet, liquid,capsule, powder and the like. However, other routes may be desirable ornecessary, particularly if the patient suffers from nausea. Such otherroutes may include, without exception, transdermal, intraperitonial,parenteral, subcutaneous, intranasal, intrathecal, intramuscular,intravenous and intrarectal modes of delivery. Another aspect of theinvention is drawn to therapeutic compositions comprising the novelcompounds of the invention and pharmaceutically acceptable salts ofthese compounds and a pharmaceutically acceptable excipient. Such anexcipient may be a carrier or a diluent; this is usually mixed with theactive compound, or permitted to dilute or enclose the active compound.If a diluent, the carrier may be solid, semi-solid, or liquid materialthat acts as an excipient or vehicle for the active compound. Theformulations may also include wetting agents, emulsifying agents,preserving agents, sweetening agents, and/or flavoring agents. If usedas in an ophthalmic or infusion format, the formulation will usuallycontain one or more salt to influence the osmotic pressure of theformulation.

In another aspect, the invention is directed to methods for thetreatment of pain, particularly chronic pain, through the administrationof one or more of the novel or otherwise known compounds of theinvention, or of pharmaceutically acceptable salts thereof to a mammalin need thereof. As indicated above, the compound will usually beformulated in a form consistent with the desired mode of delivery.

Compounds of the invention which are immunostimulants are administeredsubject to the same basic principles as the compounds having analgesicactivity, in doses which are best determined on a case-by-case and/orspecies-by-species and, in case of humans, at times on apatient-by-patient basis. Generally speaking the effective dose will bein the range of 10 μg/kg to 200 mg/kg.

In this regard it is noted that the compounds of the threo configurationare more likely to have the analgesic activity, compounds of the erythroconfiguration are more likely to have immunostimulant activity, andamong the erythro compounds those with an S configuration at carbon 2 ofthe propionic acid moiety are likely to have stronger immuno stimulantactivity.

Synthetic Methods for Obtaining the Compounds of the Invention,Experimental

The compound of the invention can be synthesized by utilizing thesynthetic methods described in the experimental below, or suchmodifications of the below described experimental methods which willbecome readily apparent to those skilled in the art in light of thepresent disclosure.

General

¹H NMR spectra were recorded at ambient temperature with an Avance 300(Bruker) spectrometer. The compounds were analyzed by reverse phase highperformance liquid chromatography (HPLC) using a Waters AutopurificationSystem equipped with a Waters 2525 Pump, a Waters 2696 photodiode arraydetector, and a XTerra column (Part. No. 186000482, 5 μm, C18, 4.5×50mm).

The HPLC method used was a gradient of 5% solvent B to 100% in 7 min.Solvent A was H₂O with 0.05% TFA and solvent B was CH₃CN with 0.05% TFA(Method A).Melting points were measured with a Büchi B-545 melting point apparatusand were uncorrected. To isolate reaction products the solvent wereremoved by evaporation using a vacuum rotatory evaporator, the waterbath temperature not exceeding 40° C.

Absolute configuration of compounds of the invention, where applicable,can generally speaking be determined in accordance with methods known inthe state of the art, such as X-ray christallography. Compounds 203 and204 are mentioned as examples for which the absolute configurations weredetermined by X-ray christallography analysis of the corresponding(1S)-camphanylamide D-(10) camphorsulfonic acid salt. As a resultCompound 204 was assigned (2S,3R). Its enantiomer, Compound 203 wasassigned by default the (2R,3S) absolute configuration.

General Synthetic Routes

The compound of the invention can be synthesized by utilizing thesynthetic methods described in a general sense immediately below and inmore detail in the experimental section of the present application, orby such modifications of the below described experimental methods whichwill become readily apparent to those skilled in the art in light of thepresent disclosure.

A general synthetic route to the novel compounds of the invention whichare amides of substituted (+/−)-threo-3-hydroxy-2-aminopropionic acid ofthe Generalized Structure 1 is described below.

In General Structure 1, for the sake of simplicity of description R*substantially corresponds to the 5, 6, or 7 membered ring structure onthe left side of Formula 1 (as the formula is depicted in the Summaryand in the instant claims) and R** substantially corresponds to the R₁groups in Formula 1.

Thus, in accordance with General Scheme 1, methyl isocyanoacetate (orethyl isocyanoacetate available commercially) is reacted with an “amine”which includes the R** groups to provide the 2-isocyanoacetic acid amidederivative shown in the general scheme. Typical examples for the aminesused in the reaction are pyrrolidine, piperidine, azetidine, morpholine,2,5-dihydro-1H-pyrrole, dialkylamines such as diethylamine, 3-fluoro-,3,3-difluoro or 3-hydroxy substituted pyrrolidines. Specific examples ofthese “amines” abound in the experimental description. The2-isocyanoacetic acid amide derivative is then reacted in the presenceof base (such as KOH) with an “aldehyde” which includes the R* group toprovide a trans “oxazoline” with high diastereoselectivity (trans:cisratios generally >97:3) as shown in the general reaction scheme 1. Thetrans oxazoline is then treated with a strong acid, such as HCl, to openthe ring and to provide thethreo-3-substituted-3-hydroxy-2-amino-propionic acid amides (withthreo:erythro ratios generally >97:3) of the invention as shown inGeneral Scheme 1.

Compounds of Formula 1 where the amino group of formula NH(R**)₂ is aweaker nucleophile, such as indoline, thiomorpholine and the like, canbe made as illustrated in Reaction Scheme 2 for the synthesis of(±)-threo-2-amino-3-hydroxy-1-(indolin-1-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 243 and(±)-threo-2-amino-3-hydroxy-1-(thiazolidin-3-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 242.

In Reaction Scheme 2 EDCl stands for1-(3-dimethylaminopropyl)-ethylcarbodiimide hydrochloride; HOBT standsfor 1-hydroxybenzotriazole; BOC₂O stands for di-t-butyl-dicarbonate andTEA stands for triethylamine.

As it will be readily understood by those skilled in the art, for a moregeneral synthetic route, such as the one shown in Reaction Scheme 2, the4-pyridyl group can be substituted with an R* group (as defined inconnection with Scheme 1) and the indoline can be substituted with otherweak nucleophilic amines of the formula NH(R**)₂ (R** defined as inconnection with Reaction Scheme 1) to provide other compounds of Formula1 analogous to compounds 242 and 243.

Isomerically pure and/or enantiomerically pure compounds and furtherderivatives of the 3-substituted-3-hydroxy-2-amino-propionic acid amidesare obtained by separation techniques and reactions which, per se, arewell known to the synthetic chemist. The experimental section of thepresent invention abounds in examples of such separation techniques andreactions. Some of the typical separation techniques and reactions aregenerally described below.

Separation of threo and erythro isomers, when both are formed in thereactions leading to the compounds of the invention, can typically beseparated by chromatographic methods. The more abundantly formed threoisomers can also be converted into the erythro isomers by oxidizing tothe ketone level the hydroxyl group in the 3 position of the propanoicacid moiety and subsequently reducing the resulting ketone to thehydroxyl level. (See, for example, the preparation of Compound 219).

Separation of enantiomeric mixtures can be performed on Chiralpackcolumns which are well known in the art. (See, for example, thepreparation of Compound 204).

The amino function in the 2-position of the propanoic acid moiety is,generally speaking, more reactive towards acylation and carbamoylationthan the hydroxyl group in the 3 position. Therefore, acylatedderivatives of the 2-amino function can be prepared by using acylchlorides such as acetyl chloride and hexanoyl chloride. (See, Method Gand the preparation of Compound 51). Carbamate derivatives of the2-amino function can be obtained by using chloroformates, such asbenzylchloroformate. (See, for example, the preparation of Compound 58).The tertiary butyl carbamoyl function can also serve as a removableprotecting group of the 2-amino function. (see for example thepreparation of Compounds 219 and 224). When the 2-amino function of thecompounds of the invention is already acylated or bears a carbamoylgroup, then the 3-hydroxy group of the propanoic acid moiety can besubjected to acylation by reagents such as acetic anhydride. (See forexample the preparation of Compound 217).

Alkylation of the 2-amino function is readily performed by condensingthe compound bearing the 2—NH₂ group with an aldehyde to obtain a Schiffbase intermediate which can be reduced, without isolation, to providethe N-alkyl, arylalkyl or heteroaryl-alkyl compound. The proceduredescribed for preparing Compound 234 can be generalized to makecompounds of the invention where the 2-amino function bears anaryl(hydroxy)alkyl or heteroaryl(hydroxy)alkyl group.

Several compounds of the invention of Formula 2 can be obtained byderivatization of compounds of Formula 1, or by such modification of thesynthetic routes leading to compounds of Formula 1 which will becomereadily apparent to those skilled in the art in light of the presentdisclosure. For example, compounds of Formula 2 where R₉ is OH or OR₁₁and R₁₀ is alkyl can be made by using a “ketone” bearing the R₁₀ group,instead of the “aldehyde” in General Reaction Scheme 1.

Compounds of Formula 2 where the R₉ and R₁₀ groups jointly form an oxime(NOH) group can be obtained by oxidizing the 3-hydroxyl group of thepropanoic acid moiety to the ketone stage and reacting the resultingketone with hydroxylamine.

Another general synthetic route for making several compounds of Formula2 is illustrated in Synthetic Scheme 3 adapted for synthesizing(R)-2-amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride, Compound 236 of the present invention.

As it will be readily understood by those skilled in the art, for a moregeneral synthetic route, such as the one shown in Reaction ShynteticScheme 3, the 4-pyridyl group can be substituted with an R* group (asdefined in connection with Scheme 1) and the pyrrolidine can besubstituted with amines of the formula NH(R**)₂ (R** defined as inconnection with Reaction Scheme 1) to provide other compounds of Formula2 analogous to compound 236 or to its enantiomer(S)-2-amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 240.

Detailed Description of the Synthesis of Preferred Compounds(Experimental) Preparation of Compound 122-Isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098

To stirred and cooled (0° C.) methyl isocyanoacetate (96% technicalgrade, 5.0 g, 47.8 mmol) was slowly added in 0.75 h pyrrolidine (6.5 mL,78 mmol). The mixture was stirred for 1.5 h with continued cooling andthen concentrated. The resulting oil was co-evaporated twice fromCH₂Cl₂:hexane to remove residual pyrrolidine.2-Isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 was obtained as ayellow solid (6.85 g, 98% yield) and used in the next step withoutpurification.

MW: 138.17; Yield: 98%; yellow solid; Mp (° C.)=73.9.

¹H-NMR (CDCl₃, *): 1.81-2.08 (m, 4H, 2×CH₂), 3.35-3.45 (m, 2H, —NCH₂),3.50-3.60 (m, 2H, —NCH₂), 4.23 (s, 2H, CH₂CO).

trans-(4,5-Dihydro-5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04100

To a stirred and cooled (0° C.) solution of potassium hydroxide (0.43mg, 7.60 mmol) in MeOH (6.5 mL) were added successively1,4-benzodioxan-6-carboxaldehyde (1.31 g, 7.96 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (1.0 g, 6.57 mmol). Thesolution was stirred 3 h at 0° C. and then concentrated. The residue waspartitioned between EtOAc (100 mL) and water. The organic layer wascombined with 2 additional EtOAc extracts (2×100 mL), washed with brine,dried over MgSO₄, filtered and evaporated. Concentration afford to acrude product which was purified by column chromatography on silica(EtOAc) to yield, after evaporation and drying, totrans-4,5-dihydro-5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04100 as a colourless oil (1.76 g, 89% yield).

MW: 440.49; Yield: 89%; colorless oil.

¹H-NMR (CDCl₃, δ): 1.75-2.10 (m, 4H, 2×CH₂), 3.40-3.59 (m, 3H,1.5×CH₂N), 3.85-4.00 (m, 1H, 0.5×CH₂N), 4.26 (s, 4H, CH₂O), 4.59 (dd,1H, J=7.5 Hz, J=2.2 Hz, CH—N), 6.00 (d, 1H, J=7.5 Hz, CH—O), 6.75-6.90(m, 3H, ArH), 7.00 (d, 1H, J=2.2 Hz, CH═N).

trans-(4,5-Dihydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07074

To a stirred and cooled (0° C.) solution of potassium hydroxide (0.37 g,6.57 mmol) in methanol (30 mL) was added a mixture of4-methoxybenzaldehyde (0.88 mL, 7.23 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (1.0 g, 6.57 mmol). Thesolution was stirred 4 h with continued cooling and then concentrated.The residue was partitioned between ethyl acetate and water. The organiclayer was combined with additional ethyl acetate extracts, washed withaqueous sodium chloride and dried over MgSO₄. Concentration afforded acrude product as a glassy solid. Flash chromatography over silica (ethylacetate) yielded totrans-(4,5-dihydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07074 as a pale yellow solid (1.2 g, 90.5%).

MW: 274.32; Yield: 90.5%; pale yellow solid; Mp (° C.): 91.2.

R_(f): 0.30 (EtOAc).

¹H-NMR (CDCl₃, *): 1.75-2.08 (m, 4H, 2×CH₂), 3.40-3.58 (m, 3H, CH₂N),3.52 (s, 3H, CH₃O), 3.88-3.98 (m, 1H, CH₂N), 4.59 (dd, 1H, J=7.6 Hz,J=2.2 Hz, CH—N), 6.06 (d, 1H, J=7.6 Hz, CH—O), 6.90 (d, 2H, J=8.7 Hz,ArH), 7.01 (d, 1H, J=2.2 Hz, CH═N), 7.25 (d, 2H, J=8.7 Hz, ArH).

MS-ESI m/z (% rel. Int.): 275.1 ([MH]⁺, 10), 247.1 (100).

HPLC: Method A, detection UV 280 nm, SLA 07074 RT=5.2 min, peak area92%.

DL-threo-2-Amino-3-hydroxy-3-(4-methoxyphenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride SLA 07078

To a stirred solution oftrans-(4,5-dihydro-5-(4-methoxyphenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07074 (1.61 g, 5.93 mmol) in methanol (13 mL) was added hydrochloricacid (1 mL). After heating at 50° C. for 3 h the mixture reaction wasconcentrated and the resulting yellow oil was co-evaporated twice withethyl acetate before solidifying. Trituration (ethyl acetate) and dryingaffordedDL-threo-2-amino-3-hydroxy-3-(4-methoxyphenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride SLA 07078 as a white solid (1.64 g, 93%).

MW: 300.78; Yield: 93%; white Solid; Mp (° C.): 177.0.

¹H-NMR (CD₃OD, *): 1.32-1.50 (m, 1H, 0.5×CH₂), 1.50-1.88 (m, 3H,1.5×CH₂), 2.15-2.28 (m, 1H, CH₂N), 3.15-3.42 (m, 3H, 1.5×CH₂N), 3.79 (s,3H, CH₃O), 4.06 (d, 1H, J=9.2 Hz, CH—N), 4.78 (d, 1H, J=9.2 Hz, CHO),6.94 (d, 2H, J=8.5 Hz, ArH), 7.34 (d, 2H, J=8.5 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.8, 26.6, 47.2, 47.6, 55.9, 59.6, 73.9, 115.0(2×C), 128.9 (2×C), 132.5, 161.7, 166.4.

DL-threo-2-amino-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 12

To a stirred solution oftrans-4,5-dihydro-5-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04100 (1.74 g, 5.77 mmol) in methanol (15 mL) was added hydrochloricacid (1 mL). After heating at 50° C. for 3 h the mixture reaction wasconcentrated and the resulting yellow oil was co-evaporated twice withethyl acetate before solidifying. Trituration (ethyl acetate) and dryingaffordedDL-threo-2-amino-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 12 as a white solid (1.85 g, 95%).

MW: 328.79; Yield: 95.0%; White Solid; Mp (° C.): 176.2.

¹H-NMR (CD₃OD, *): 1.42-1.58 (m, 1H, 0.5×CH₂), 1.58-1.70 (m, 1H,0.5×CH₂), 1.70-1.88 (m, 2H, CH₂), 3.20-3.45 (m, 4H, 2×N—CH₂), 4.06 (d,1H, J=9.1 Hz, CH—N), 4.25 (s, 2H, OCH₂), 4.75 (d, 1H, J=9.2 Hz, CH—O),4.89 (s, 2H, OCH₂), 6.82-6.95 (m, 3H, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 26.7, 47.3, 47.6, 59.5, 65.7, 73.6, 116.4,118.3, 120.3, 133.7, 145.1, 145.6, 166.4.

Preparation of Compound 18. Method B:

To a stirred and cooled (0° C.) solution of potassium hydroxide (380 mg,5.80 mmol) in MeOH (5 mL) were added successively aldehyde (5.80 mmol)and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.8 g, 5.8 mmol).The solution was stirred 3 h at 0° C. and then concentrated. The residuewas partitioned between CH₂Cl₂ (100 mL) and water. The organic layer waswashed with brine, dried over MgSO₄, filtered and evaporated.Concentration afford to a crude product which was purified by columnchromatography on silica (cyclohexane:EtOAc=70:30 to 0:100) to yield,after evaporation and drying, to an intermediate oxazoline. To a stirredsolution of oxazoline in methanol (15 mL) was added hydrochloric acid (1mL, 12 mmol). After heating at 60° C. for 2 h, the mixture reaction wasthen concentrated and the resulting yellow oil was coevaporated twicewith MeOH before solidifying. Trituration in EtOAc:MeOH=10:1 followed byfiltration gave title compound as a white solid.

DL-threo-2-Amino-3-(biphenyl-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 18

The compound was prepared according to method B with4-phenylbenzaldehyde (1.05 g, 5.78 mmol).DL-threo-2-Amino-3-(biphenyl-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 18 was obtained as a pale brown solid (0.55 g,28% yield).

MW: 346.85; Yield: 28%; Pale Brown Solid; Mp (° C.): 197.3.

¹H-NMR (CD₃OD, *): 1.25-1.42 (m, 1H, 0.5×CH₂), 1.50-1.60 (m, 1H,0.5×CH₂), 1.60-1.80 (m, 1H, 0.5×CH₂), 2.20-2.30 (m, 2H, N—CH₂),3.15-3.30 (m, 2H, N—CH₂), 3.30-3.45 (1H, m, N—CH₂), 4.13 (d, 1H, J=9.2Hz, CH—N), 4.85-4.95 (m, 1H, CH—O), 7.32-7.38 (m, 1H, ArH), 7.46 (dd,2H, J=7.1 Hz, J=7.8 Hz, ArH), 7.52 (d, 2H, J=8.3 Hz, ArH), 7.58-7.70 (m,4H, ArH).

¹³C-NMR (CD₃OD, *): 24.8, 26.5, 47.2, 47.6, 59.5, 78.7, 127.9, 128.1,128.2, 128.8, 130.0, 139.7, 141.6, 143.3, 166.3.

MS-ESI m/z (% rel. Int.): 311.2 ([MH]⁺, 60).

HPLC: Method A, detection UV 254 nm, Compound 18 RT=4.50 min, peak area99.9%.

Preparation of 2-isocyano derivatives: SLA 07116B SLA 07116C SLA 07118,SLA 07130A, SLA 07178 and SLA 07184A.

2-Isocyano-1-(piperidin-1-yl)ethanone SLA 07116B

Prepared in accordance with Method B with methyl isocyanoacetate (2.46g, 24.63 mmol) and piperidine (3.22 mL, 37.85 mmol). The reactionmixture was stirred 1 h at RT and then concentrated. The residue wasdissolved in dichloromethane (50 mL) and the organic layer was washedwith 10% aqueous citric acid (2×25 mL), dried over MgSO₄, filtered andevaporated. 2-Isocyano-1-(piperidin-1-yl)ethanone SLA 07116B wasobtained as an orange solid (3.13 g, 83% yield).

MW: 152.19; Yield: 83%; Orange Solid; Mp (° C.): 81.6.

¹H-NMR (CDCl₃, *): 1.56-1.74 (m, 6H, CH₂C), 3.33 (t, 2H, J=5.7 Hz,CH₂N), 3.58 (t, 2H, J=5.7 Hz, CH₂N), 4.29 (s, 2H, CH₂CO).

tert-Butyl 4-(2-isocyanoacetyl)piperazine-1-carboxylate SLA 07116C

Prepared in accordance with Method B with methyl isocyanoacetate (2.51g, 25.29 mmol) and piperazine-1-carboxylic acid tert-butyl ester (6.28g, 33.85 mmol. The reaction mixture was stirred 1 h at RT and thenconcentrated. The residue was dissolved in dichloromethane (50 mL) andthe organic layer was washed with 10% aqueous citric acid (2×25 mL),dried over MgSO₄, filtered and evaporated. tert-Butyl4-(2-isocyanoacetyl)piperazine-1-carboxylate SLA 07116C was obtained asa colorless oil (0.41 g, 6.5% yield).

MW: 253.14; Yield: 6.5%; Colorless oil.

¹H-NMR (CDCl₃, *): 1.47 (s, 9H, tBu), 3.38 (t, 2H, J=5.3 Hz, CH₂N),3.45-3.53 (m, 4H, CH₂N), 3.62 (t, 2H, J=5.5 Hz, CH₂N), 4.32 (s, 2H,CH₂CO).

2-Isocyano-1-morpholinoethanone SLA 07118

Prepared in accordance with Method B with methyl isocyanoacetate (2.51g, 25.30 mmol) and morpholine (3.30 mL, 38.05 mmol). The reactionmixture was stirred 24 h at RT and then concentrated. The residue wasdissolved in dichloromethane (50 mL) and the organic layer was washedwith 10% aqueous citric acid (2×25 mL), dried over MgSO₄, filtered andevaporated. 2-Isocyano-1-morpholinoethanone SLA 07118 was obtained as abrown oil (2.28 g, 58% yield).

MW: 154.17; Yield: 58%; Brown Oil.

R_(f): 0.20 (EtOAc:cyclohexane=50:50).

¹H-NMR (CDCl₃, *): 3.42 (t, 2H, J=4.9 Hz, CH₂N), 3.65 (t, 2H, J=5.1 Hz,CH₂N), 3.73 (t, 4H, J=5.0 Hz, CH₂O), 4.31 (s, 2H, CH₂CO).

2-Isocyano-1-thiomorpholinoethanone SLA 07130A

Prepared in accordance with Method B with methyl isocyanoacetate (2.50g, 25.28 mmol) and thiomorpholine (4.25 mL, 37.85 mmol). The reactionmixture was stirred 22 h at RT and then concentrated. The residue wasdissolved in dichloromethane (50 mL) and the organic layer was washedwith 10% aqueous citric acid (2×25 mL), dried over MgSO₄, filtered andevaporated. 2-Isocyano-1-thiomorpholinoethanone SLA 07130A was obtainedas a yellow solid (3.05 g, 71% yield).

MW: 170.23; Yield: 71%; Yellow Solid; Mp (° C.): 144.4.

R_(f): 0.35 (EtOAc:cyclohexane=50:50).

¹H-NMR (CDCl₃, *): 2.68 (m, 4H, 2×CH₂S), 3.67 (m, 2H, N—CH₂), 3.90 (m,2H, N—CH₂), 4.31 (s, 2H, COCH₂).

2-Isocyano-1-(2H-pyrrol-1(5H)-yl)ethanone SLA 07178

Prepared in accordance with Method B with methyl isocyanoacetate (1.00g, 10.10 mmol) and dihydro-1H-pyrrole (1.01 mL, 15.15 mmol). Thereaction mixture was stirred 5 h at 50° C. and concentrated. The residuewas dissolved in dichloromethane (50 mL) and the organic layer waswashed with 10% aqueous citric acid (2×25 mL), dried over MgSO₄,filtered and evaporated. 2-Isocyano-1-(2H-pyrrol-1(5H)-yl)ethanone SLA07178 was obtained (1.0 g, 73% yield) as a yellow solid.

MW: 136.15; Yield: 73%; Yellow Solid.

R_(f): 0.35 (EtOAc:cyclohexane=50:50).

¹H-NMR (CDCl₃, *): 4.23 (s, 4H, 2×CH₂N), 4.31 (s, 2H, CH₂N), 5.80-5.86(m, 1H, CH═C), 5.90-5.95 (m, 1H, CH═C).

N,N-Diethyl-2-isocyanoacetamide SLA 07184A

Prepared in accordance with Method B with methyl isocyanoacetate (2.50g, 25.29 mmol) and diethylamine (1.96 mL, 37.94 mmol). The reactionmixture was stirred 5 h at 50° C. and concentrated. The residue wasdissolved in dichloromethane (50 mL) and the organic layer was washedwith 10% aqueous citric acid (2×25 mL), dried over MgSO₄, filtered andevaporated. N,N-Diethyl-2-isocyanoacetamide SLA 07184A was obtained(1.213 g, 34% yield) as a brown oil.

MW: 140.18; Yield: 34%; Brown Oil.

R_(f): 0.35 (EtOAc:cyclohexane=50:50).

¹H-NMR (CDCl₃, *): 1.15-1.26 (m, 6H, CH₃), 3.21-3.30 (m, 2H, CH₂N),3.38-3.45 (m, 2H, CH₂N), 4.26 (s, 2H, CH₂CO).

Preparation of Oxazolines: BLE 04110B, SLA 07122A, SLA 07124A, SLA07124B, SLA 07132, BLE 04110A, Compound 19, BLE 04124A, BLE 04124B, BLE04124C, BLE 04124D, BLE 04130B, BLE 04130C, BLE 04130D, BLE 04136B, BLE04136C, BAL 01016, BLE 04136D, BAL 01014, SLA 07194A, SLA 07174, BAL01028A, BLA 01028B, SLA 07158 and SLA 07180.

trans-(4,5-Dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110B General Method D for Oxazolines Formation:

To a stirred and cooled (0° C.) solution of potassium hydroxide (0.55 g,9.80 mmol) in methanol (10 mL) were added a mixture of 3-pyridinecarboxaldehyde (1.03 mL, 10.84 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (1.50 g, 10.86 mmol).The solution was stirred 3 h at 0° C. and then concentrated. The residuewas partitioned between ethyl acetate (100 mL) and water. The organiclayer was combined with two additional ethyl acetate extracts (2×100mL), washed with aqueous sodium chloride and dried over MgSO₄, filteredand evaporated. Concentration afforded a crude product which waspurified by column chromatography on silica (CH₂Cl₂:MeOH=98:2) to yieldtotrans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110B (0.95 g, 39% yield) as a pale yellow pale solid.

MW: 245.28; Yield: 39%; Yellow Pale Solid; Mp (° C.): 107.0.

¹H-NMR (CDCl₃, *): 1.78-2.10 (m, 4H, 2×CH₂), 3.40-3.61 (m, 3H, CH₂N),3.90-4.04 (m, 1H, CH₂N), 4.59 (dd, 1H, J=7.7 Hz, J=2.2 Hz, CH—N), 6.21(d, 1H, J=7.7 Hz, CH—O), 7.04 (d, 1H, J=2.2 Hz, O—CH═N), 7.33 (m, 1H,ArH), 7.64 (m, 1H, ArH), 8.59 (d, 2H, J=2.8 Hz, ArH).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.4, 46.6, 75.7, 79.3, 123.7, 133.5,135.3, 147.6, 149.9, 155.2, 166.2.

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(piperidin-1-yl)methanoneSLA 07122A

SLA 07122A was prepared in accordance with method method D using2-isocyano-1-(piperidin-1-yl)ethanone (0.4 g, 26.3 mmol), potassiumhydroxide (0.15 g, 26.7 mmol) in methanol (5 mL) andpyridine-4-carbaldehyde (0.37 mL, 40.9 mmol). The solution was stirred20 h at 0° C.trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(piperidin-1-yl)methanoneSLA 07122A was obtained as a yellow solid (0.353 g, 52% yield).

MW: 259.30; Yield: 52%; Yellow Solid; Mp (° C.): 111.7.

R_(f): 0.80 (MeOH:CH₂Cl₂=10:90).

¹H-NMR (CDCl₃, *): 1.55-1.78 (m, 6H, 3×CH₂), 3.45-3.60 (m, 2H, CH₂N),3.70-3.85 (m, 2H, CH₂N), 4.60 (dd, 1H, J=7.8 Hz, J=2.3 Hz, CH—N), 6.27(d, 1H, J=7.8 Hz, CH—O), 7.01 (d, 1H, J=2.3 Hz, CH═N), 7.23 (dd, 2H,J=4.5 Hz, J=1.6 Hz, ArH), 8.61 (dd, 2H, J=4.5 Hz, J=1.5 Hz, ArH).

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(morpholino)methanone SLA07124A

SLA 07118 was prepared in accordance with method D using2-isocyano-1-morpholinoethanone (0.40 g, 25.95 mmol), potassiumhydroxide (0.146 g, 26.0 mmol) in methanol (5 mL) andpyridine-4-carbaldehyde (0.36 mL, 40.4 mmol). The solution was stirred22 h at 0° C.trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(morpholino)methanone SLA07124A was obtained as a yellow solid (0.168 g, 25% yield).

MW: 261.28; Yield: 25%; Yellow Solid; Mp (° C.): 90.5.

R_(f): 0.30 (EtOAc:cyclohexane=20:80).

¹H-NMR (CDCl₃, *): 3.46-4.02 (m, 8H, 2×CH₂O, 2×CH₂N), 4.56 (dd, 1H,J=7.8 Hz, J=2.3 Hz, CH—N), 6.27 (d, 1H, J=7.9 Hz, CH—O), 7.02 (d, 1H,J=2.3 Hz, CH═N), 7.24 (dd, 2H, J=4.6 Hz, J=1.4 Hz, ArH), 8.63 (dd, 2H,J=4.5 Hz, J=1.6 Hz, ArH).

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-tert-butyloxycarbonylpiperazin-1-yl)methanoneSLA 07124B

SLA 07124B was prepared in accordance with method D using tert-butyl4-(2-isocyanoacetyl)piperazine-1-carboxylate SLA 07116C (0.41 g, 16.20mmol), potassium hydroxide (0.91 g, 16.2 mmol) in methanol (5 mL) andpyridine-4-carbaldehyde (0.227 mL, 25.2 mmol). The solution was stirred22 h at 0° C.trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-tert-butyloxycarbonyl-piperazin-1-yl)methanoneSLA 07124B was obtained as a pale yellow solid (0.335 g, 58% yield).

MW: 360.41; Yield: 58%; Pale Yellow Solid; Mp (° C.): 157.2° C.

¹H-NMR (CDCl₃, *): 1.47 (s, 9H, tBu), 3.25-4.02 (m, 8H, CH₂N), 4.58 (dd,1H, J=7.8 Hz, J=2.3 Hz, CH—N), 6.27 (d, 1H, J=7.8 Hz, CH—O), 7.01 (d,1H, J=2.3 Hz, CH═N), 7.24 (dd, 2H, J=4.6 Hz, J=1.4 Hz, ArH), 8.62 (dd,2H, J=4.5 Hz, J=1.6 Hz, ArH).

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(thiomorpholino)methanoneSLA 07132

SLA 07132 was prepared in accordance with method D using2-Isocyano-1-thiomorpholinoethanone SLA 07130A (0.752 g, 4.41 mmol),potassium hydroxide (0.250 g, 4.45 mmol) in methanol (10 mL) andpyridine-4-carbaldehyde (0.436 mL, 4.85 mmol). The solution was stirred24 h at 0° C.trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(thiomorpholino)methanoneSLA 07132 was obtained as a yellow foam (1.01 g, 83%).

MW: 277.35; Yield: 83%; Yellow Foam.

R_(f): 0.80 (MeOH:CH₂Cl₂=10:90).

¹H-NMR (CDCl₃, *): 2.53-2.92 (m, 4H, 2×CH₂), 3.58-3.70 (m, 1H, CH₂N),3.78-3.88 (m, 1H, CH₂N), 4.15-4.30 (m, 2H, CH₂N), 4.56 (dd, J=7.8 Hz,J=2.3 Hz, 2H, CH—N), 6.27 (d, 1H, J=7.8 Hz, CH—O), 7.02 (d, 1H, J=2.3Hz, N═CH—O), 7.22 (d, 2H, J=6.1 Hz, ArH), 8.61 (dd, 2H, J=6.1 Hz, ArH).

¹³C-NMR (CDCl₃, *): 27.3, 28.0, 45.4, 48.6, 74.9, 79.6, 120.0 (2×C),148.5, 150.3 (2×C), 154.8, 166.2.

trans-(4,5-Dihydro-5-(pyridin-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110A

BLE 04110A was prepared in accordance with method D using 2-pyridinecarboxaldehyde (1.02 mL, 10.84 mmol).Trans-(4,5-dihydro-5-(pyridin-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110A was obtained as a yellow pale oil (0.45 g, 19% yield).

MW: 245.28; Yield: 19%; Yellow Pale Oil.

¹H-NMR (CDCl₃, *): 1.73-2.08 (m, 4H, 2×CH₂), 3.35-3.70 (m, 3H, CH₂N),3.85-4.00 (m, 1H, CH₂N), 5.05 (dd, 1H, J=6.9 Hz, J=2.2 Hz, CH—N), 6.18(d, 1H, J=6.9 Hz, CH—O), 7.02 (d, 1H, J=2.1 Hz, O—CH═N), 7.25 (m, 1H,ArH), 7.43 (d, 1H, J=7.8 Hz, ArH), 7.69 (dt, 1H, J=7.8 Hz, J=1.8 HzArH), 8.62 (m, 1H, ArH).

¹³C-NMR (CDCl₃, *): 24.2, 25.9, 46.3, 46.5, 73.4, 81.3, 121.5, 123.2,136.8, 149.8, 154.8, 158.0, 166.9.

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneCompound 19

SLA 07092 was prepared in accordance with method D usingpyridine-4-carbaldehyde (1.88 mL, 19.76 mmol), KOH (1.01 g, 18.00 mmol)in methanol (18 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098(2.73 g, 19.76 mmol). The residue was partitioned between ethyl acetate(200 mL) and water (150 mL). The organic layer was combined withadditional ethyl acetate extracts (2×150 mL), washed with aqueous sodiumchloride (2×150 mL) and dried over MgSO₄, filtered and evaporated.Trans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneCompound 19 was obtained as a white solid (4.32 g, 98% yield).

MW: 245.28; Yield: 98%; White Solid; Mp (° C.)=69.2.

R_(f): 0.65 (MeOH:CH₂Cl₂=10:90).

¹H-NMR (CDCl₃, *): 1.78-2.06 (m, 4H, 2×CH₂), 3.44-3.60 (m, 3H, CH₂N),3.90-4.01 (m, 1H, CH₂N), 4.52 (dd, 1H, J=7.9 Hz, J=2.2 Hz, CH—N), 6.19(d, J=7.9 Hz, 1H, CH—O), 7.03 (d, 1H, J=2.2 Hz, N═CH—O), 7.24 (dd, 2H,J=4.5 Hz, J=1.5 Hz, ArH), 8.61 (dd, 2H, J=4.5 Hz, J=1.5 Hz, ArH).

trans-(4,5-Dihydro-5-(thiophen-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124A

BLE 04124A was prepared in accordance with method D usingthiophen-3-carboxaldehyde (0.475 mL, 5.42 mmol), KOH (0.276 mg, 4.92mmol) in methanol (5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE04098 (0.75 g, 5.43 mmol). After work-up the residue obtained wasrecristallized from ethyl acetate to obtain after filtrationtrans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110A as a yellow pale solid (0.498 g, 40% yield).

MW: 250.32; Yield: 40.5%; Yellow Pale Solid; Mp (° C.): 105.9.

¹H-NMR (CDCl₃, *): 1.78-2.10 (m, 4H, CH₂), 3.42-3.61 (m, 3H, CH₂N),3.90-4.02 (m, 1H, CH₂N), 4.63 (dd, 1H, J=7.4 Hz, J=2.2 Hz, CH—N), 6.20(d, 1H, J=7.4 Hz, CH—O), 6.98 (d, 1H, J=2.2 Hz, O—CH═N), 7.03 (dd, 1H,J=5.0 Hz, J=1.3 Hz, CH═C), 7.30 (dt, 1H, J=3.0 Hz, J=1.3 Hz, CH═C), 7.36(dd, 1H, J=5.0 Hz, J=3.0 Hz, CH═C).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.4, 46.6, 74.6, 77.9, 122.7, 125.1,127.3, 140.4, 155.3, 166.7.

MS-ESI m/z (% rel. Int.): 251.0 ([MH]⁺, 17), 223 (40), 179.9 (60), 151.9(63), 123.9 (100).

HPLC: Method A, detection UV 254 nm, BLE 04124A, RT=4.4 min, peak area98.0%.

trans-(4,5-Dihydro-5-(thiophen-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124B

BLE 04124B was prepared in accordance with method D usingthiophen-2-carboxaldehyde (0.507 mL, 5.42 mmol), KOH (0.276 mg, 4.92mmol) in methanol (5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE04098 (0.75 g, 5.43 mmol). After work-up the residue obtained waspurified by column chromatography (EtOAc) to led after evaporation totrans-(4,5-dihydro-5-(thiophen-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124B as a yellow pale solid (0.713 g, 58% yield).

MW: 250.32; Yield: 58%; Yellow Pale Solid; Mp (° C.): 71.3.

¹H-NMR (CDCl₃, *): 1.78-2.10 (m, 4H, CH₂), 3.42-3.62 (m, 3H, CH₂N),3.90-4.03 (m, 1H, CH₂N), 4.76 (dd, 1H, J=7.3 Hz, J=2.2 Hz, CH—N), 6.37(d, 1H, J=7.3 Hz, CH—O), 6.96 (d, 1H, J=2.2 Hz, O—CH═N), 7.00 (dd, 1H,J=5.0 Hz, J=3.5 Hz, CH═C), 7.11 (d, 1H, J=3.1 Hz, CH═C), 7.33 (dd, 1H,J=5.0 Hz, J=0.7 Hz, CH═C).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.4, 46.6, 75.5, 77.6, 126.3 (2×C),127.1, 142.0, 154.9, 166.3.

MS-ESI m/z (% rel. Int.): 251.0 ([MH]⁺, 15), 223 (100).

HPLC: Method A, detection UV 254 nm, BLE 04124B, RT=3.8 min, peak area>90%.

trans-(4,5-Dihydro-5-(thiazol-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124C

BLE 04124C was prepared in accordance with method D using2-thiazolecarboxaldehyde (0.476 mL, 5.42 mmol), KOH (0.276 mg, 4.92mmol) in methanol (5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE04098 (0.75 g, 5.43 mmol). After work-up the residue obtained waspurified by column chromatography (EtOAc) to led after evaporation totrans-(4,5-dihydro-5-(thiazol-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124C as a colourless oil (0.564 g, 45.5% yield).

MW: 251.3; Yield: 45.5%; colourless Oil.

¹H-NMR (CDCl₃, *): 1.80-2.10 (m, 4H, CH₂), 3.47-3.70 (m, 3H, CH₂N),3.91-4.02 (m, 1H, CH₂N), 5.18 (dd, 1H, J=6.4 Hz, J=2.2 Hz, CH—N), 6.40(d, 1H, J=6.4 Hz, CH—O), 6.97 (d, 1H, J=2.2 Hz, O—CH═N), 7.38 (d, 1H,J=3.3 Hz, CH═C), 7.81 (d, 1H, J=3.3 Hz, CH═C).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.4, 46.5, 73.7, 78.2, 120.1, 143.3,154.3, 166.1, 168.2.

MS-ESI m/z (% rel. Int.): 252.0 ([MH]⁺, 18), 225 (30), 198.9 (37), 153.9(48), 143.0 (100).

HPLC: Method A, detection UV 254 nm, BLE 04124C, RT=3.5 min, peak area>90%.

trans-(5-(Benzo[b]thiophen-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124D

BLE 04124D was prepared in accordance with method D usingthianaphtene-3-carboxaldehyde (0.88 g, 5.42 mmol), KOH (0.276 mg, 4.92mmol) in methanol (5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE04098 (0.75 g, 5.43 mmol). After work-up the residue obtained waspurified by column chromatography (EtOAc) to led after evaporation totrans-(5-(benzo[b]thiophen-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124D as a white solid (1.12 g, 75.5% yield).

MW: 300.38; Yield: 75.5%; White Solid; Mp (° C.): 92.2.

¹H-NMR (CDCl₃, *): 1.75-2.08 (m, 4H, CH₂), 3.36-3.49 (m, 1H, CH₂N),3.50-3.62 (m, 1H, CH₂N), 3.89-4.00 (m, 1H, CH₂N), 4.75 (dd, 1H, J=7.6Hz, J=2.2 Hz, CH—N), 6.54 (d, 1H, J=7.6 Hz, CH—O), 7.08 (d, 1H, J=2.2Hz, O—CH═N), 7.35 (m, 2H, ArH), 7.45 (s, 1H, C═CH—S), 7.67-7.75 (m, 1H,ArH), 7.84-7.92 (m, 1H, ArH).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.5, 46.6, 73.3, 77.7, 121.8, 123.1,124.1, 124.6, 124.8, 134.0, 136.4, 141.0, 155.4, 166.6.

MS-ESI m/z (% rel. Int.): 301.0 ([MH]⁺, 30), 273.0 (100).

HPLC: Method A, detection UV 254 nm, BLE 04124D, RT=4.2 min, peak area92.0%.

trans-(5-(Furan-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130B

BLE 04130B was prepared in accordance with method D using 3-furaldehyde(0.453 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol).After work-up the residue was washed with a minimum of ethyl acetate toled, after filtration and drying, totrans-(5-(furan-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130B as a white solid (0.837 g, 72.5% yield).

MW: 234.25; Yield: 72.5%; White Solid; Mp (° C.): 136.7.

¹H-NMR (CDCl₃, *): 1.80-2.10 (m, 4H, CH₂), 3.47-3.58 (m, 3H, CH₂N),3.91-4.02 (m, 1H, CH₂N), 4.61 (dd, 1H, J=7.3 Hz, J=2.1 Hz, CH—N), 6.10(d, 1H, J=7.3 Hz, CH—O), 6.36 (dd, 1H, J=1.6 Hz, J=0.6 Hz, CH═C), 6.95(d, 1H, J=2.1 Hz, O—CH═N), 7.44 (t, 1H, J=1.6 Hz, OCH═C); 7.50 (d, 1H,J=0.6 Hz, OCH═C).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.4, 46.6, 70.0, 74.0, 108.1, 124.0,140.4, 144.2, 155.3, 166.6.

trans-(4,5-Dihydro-5-(naphthalen-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130C

BLE 04130C was prepared in accordance with method D using2-naphtaldehyde (0.847 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) inmethanol (5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098(0.75 g, 5.43 mmol). After work-up the residue was washed with a minimumof ethyl acetate to led, after filtration and drying, totrans-(4,5-dihydro-5-(naphthalen-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130C as a white solid (0.791 g, 54.5% yield).

MW: 294.35; Yield: 54.5%; White Solid; Mp (° C.): 117.9.

¹H-NMR (CDCl₃, *): 1.78-2.07 (m, 4H, CH₂), 3.37-3.49 (m, 1H, CH₂N),3.49-3.61 (m, 2H, CH₂N), 3.88-3.99 (m, 1H, CH₂N), 4.67 (dd, 1H, J=7.7Hz, J=2.2 Hz, CH—N), 6.31 (d, 1H, J=7.7 Hz, CH—O), 7.10 (d, 1H, J=2.2Hz, O—CH═N), 7.38 (dd, 1H, J=8.5 Hz, J=1.7 Hz, ArH); 7.45-7.54 (m, 2H,ArH), 7.79-7.90 (m, 4H, ArH).

¹³C-NMR (CDCl₃, *): 23.8, 25.7, 46.1, 46.2, 75.3, 81.4, 122.7, 124.9,126.1, 126.2, 127.4, 127.7, 128.7, 132.8, 132.9, 136.5, 155.2, 166.4.

MS-ESI m/z (% rel. Int.): 295.1 ([MH]⁺, 40), 267.1 (100).

HPLC: Method A, detection UV 254 nm, BLE 04130C, RT=4.2 min, peak area92.0%.

trans-(4,5-Dihydro-5-(naphthalen-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130D

BLE 04130D was prepared in accordance with method D using1-naphtaldehyde (0.736 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) inmethanol (5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098(0.75 g, 5.43 mmol). After work-up the residue was purified by columnchromatography on silica (EtOAc:cyclohexane=80:20 to 90:10) to led,after evaporation, totrans-(4,5-dihydro-5-(naphthalen-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130D as a colorless gum (0.850 g, 58.5% yield).

MW: 294.35; Yield: 58.5%; Colorless gum.

¹H-NMR (CDCl₃, *): 1.75-2.02 (m, 4H, CH₂), 3.25-3.37 (m, 1H, CH₂N),3.52-3.67 (m, 2H, CH₂N), 3.82-3.93 (m, 1H, CH₂N), 4.62 (dd, 1H, J=7.0Hz, J=2.0 Hz, CH—N), 6.89 (d, 1H, J=7.0 Hz, CH—O), 7.16 (d, 1H, J=2.0Hz, O—CH═N), 7.44-7.58 (m, 4H, ArH), 7.80-7.90 (m, 3H, ArH).

¹³C-NMR (CDCl₃, *): 24.2, 25.9, 46.5 (2×C), 75.3, 79.2, 122.5, 123.0,125.4, 126.0, 126.8, 128.7, 129.0, 129.9, 133.9, 135.5, 155.5, 166.9.

MS-ESI m/z (% rel. Int.): 295.1 ([MH]⁺, 50), 267.1 (100).

HPLC: Method A, detection UV 254 nm, BLE 04130D, RT=4.2 min, peak area95.0%.

trans-(4,5-Dihydro-5-(quinolin-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136B

BLE 04136B was prepared in accordance with method D using 2-quinolinecarbaldehyde (0.852 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol(5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.75 g,5.43 mmol). After work-up the residue was purified by columnchromatography on silica (EtOAc) to led, after evaporation, totrans-(4,5-dihydro-5-(quinolin-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136B as a yellow pale solid (0.966 g, 60.3% yield).

MW: 295.34; Yield: 60.3%; Yellow Pale Solid; Mp (° C.): 93.8.

¹H-NMR (CDCl₃, *): 1.85-2.10 (m, 4H, CH₂), 3.50-3.66 (m, 2H, CH₂N),3.67-3.80 (m, 1H, CH₂N), 3.92-4.03 (m, 1H, CH₂N), 5.32 (dd, 1H, J=7.8Hz, J=2.1 Hz, CH—N), 6.31 (d, 1H, J=7.8 Hz, CH—O), 7.06 (d, 1H, J=2.1Hz, O—CH═N), 7.51-7.60 (m, 2H, ArH); 7.72 (t, 1H, J=8.4 Hz, ArH), 7.83(t, 1H, J=8.1 Hz, ArH), 8.07 (d, 1H, J=8.4 Hz, ArH), 8.20 (d, 1H, J=8.4Hz, ArH).

¹³C-NMR (CDCl₃, *): 24.3, 26.2, 46.5, 46.7, 73.0, 82.1, 119.2, 126.9,127.7, 127.8, 129.5, 129.9, 137.2, 147.7, 155.0, 158.2, 167.3.

trans-(4,5-Dihydro-5-(isoquinolin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136C

BLE 04136C was prepared in accordance with method D using 4-quinolinecarbaldehyde (0.852 g, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol(5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.75 g,5.43 mmol). After work-up the residue was washed with a minimum of EtOActo led, after filtration and drying, totrans-(4,5-dihydro-5-(isoquinolin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136C as a white solid (0.640 g, 40% yield).

MW: 295.34; Yield: %; White Solid; Mp (° C.): 152.0.

¹H-NMR (CDCl₃, *): 1.85-2.08 (m, 4H, CH₂), 3.28-3.40 (m, 1H, CH₂N),3.54-3.69 (m, 2H, CH₂N), 3.84-3.95 (m, 1H, CH₂N), 4.57 (dd, 1H, J=6.8Hz, J=2.1 Hz, CH—N), 6.93 (d, 1H, J=6.8 Hz, CH—O), 7.15 (d, 1H, J=2.1Hz, O—CH═N), 7.41 (d, 1H, J=4.5 Hz, ArH); 7.59 (m, 1H, ArH), 7.76 (m,1H, ArH), 7.91 (d, 1H, J=8.3 Hz, ArH), 8.16 (d, 1H, J=8.3 Hz, ArH), 8.92(d, 1H, J=4.5 Hz, ArH).

¹³C-NMR (CDCl₃, *): 24.2, 25.9, 46.7 (2×C), 75.6, 77.8, 116.4, 123.1,124.8, 127.5, 129.6, 130.5, 145.5, 148.4, 150.3, 155.0, 166.0.

trans-(4,5-Dihydro-5-(quinolin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01016

To a stirred and cooled (0° C.) solution of KOH (0.31 g, 5.43 mmol) in 5mL MeOH were added successively quinoline-3-carboxaldehyde (0.85 g, 5.43mmol) and 2-isocyano-1-pyrrolidin-1-yl-ethanone BLE 04134 (0.75 g, 5.43mmol). The mixture was stirred at 0° C. until precipitation andconcentrated. The mixture was partitioned between EtOAc (50 mL) and H₂O(25 mL). The aqueous layer was extracted twice with EtOAc (25 ml). TheEtOAc fractions were combined, washed twice with brine (2×25 mL), driedover MgSO₄ and filtered. After evaporation and dryingtrans-(4,5-dihydro-5-(quinolin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01016 was obtained (0.96 g, 60% yield) as a white solid.

MW: 295.34; Yield: 60%; White Solid; Mp (° C.): 144.4.

Rf: 0.15 (EtOAc).

¹H NMR (CDCl₃, *): 1.75-2.10 (m, 4H, 2×CH₂), 3.40-3.62 (m, 3H, CH₂N),3.90-4.05 (m, 1H, CH₂N), 4.70 (dd, 1H, J=7.8 Hz, J=2.2 Hz, CH—N), 6.40(d, 1H, J=7.8 Hz, CH—O), 7.10 (d, 1H, J=2.2 Hz, OCH═N), 7.58 (dt, 1H,J=1.1 Hz, J=8.0 Hz, ArH)), 7.73 (dt, 1H, J=1.4 Hz, J=6.9 Hz, ArH), 7.83(dd, 1H, J=1.2 Hz, J=8.2 Hz, ArH), 8.12 (m, 2H, ArH), 8.87 (d, 1H, J=2.2Hz, ArH).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.6, 46.6, 75.8, 79.7, 127.3, 127.5,127.9, 129.4, 130.0, 132.3, 133.2, 148.1, 148.4, 155.3, 166.2.

MS-ESI m/z (% rel. Int.): 296.1 ([MH]⁺, 5), 314.1 (100).

trans-(5-(Furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136D

BLE 04136C was prepared in accordance with method D using 2-furaldehyde(0.449 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.75 g, 5.43 mmol).After work-up the residue was purified by column chromatography onsilica (cyclohexane:EtOAc=100:0 to 0:100) to led, after evaporation, totrans-(5-(furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136D as a yellow pale oil (0.742 g, 58.5% yield).

MW: 234.25; Yield: 58.5%; Yellow Pale Oil.

¹H-NMR (CDCl₃, *): 1.80-2.10 (m, 4H, CH₂), 3.47-3.60 (m, 3H, CH₂N),3.94-4.06 (m, 1H, CH₂N), 4.94 (dd, 1H, J=7.4 Hz, J=2.2 Hz, CH—N), 6.14(d, 1H, J=7.4 Hz, CH—O), 6.37 (dd, 1H, J=3.3 Hz, J=1.8 Hz, CH═C), 6.48(d, 1H, J=3.3 Hz, CH═C), 6.93 (d, 1H, J=2.2 Hz, O—CH═N), 7.44 (d, 1H,J=1.8 Hz, OCH═C).

¹³C-NMR (CDCl₃, *): 24.2, 26.0, 46.4, 46.5, 71.3, 74.5, 110.2, 110.5,143.6, 150.4, 155.0, 166.3.

trans-(4,5-Dihydro-5-(2-methoxypyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01014

BAL 01014 was prepared in accordance with method D using2-methoxy-3-pyridinecarboxaldehyde (0.64 ml, 5.43 mmol), KOH (0.305 mg,5.43 mmol) in methanol (5 mL) and 2-isocyano-1-(pyrrolidin-1-yl)ethanoneBLE 04098 (0.75 g, 5.43 mmol). After work-uptrans-(4,5-dihydro-5-(2-methoxypyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01014 was obtained (0.74 mg, 50% yield) as a white solid.

MW: 275.30; Yield: 50%; White Solid; Mp (° C.): 110.1.

Rf: 0.25 (EtOAc).

¹H NMR (CDCl₃, *): 1.82-2.10 (m, 4H, 2×CH₂), 3.40-3.62 (m, 3H, CH₂N),3.80-3.90 (m, 3H, CH₂N), 3.93 (s, 3H, OMe), 4.61 (dd, 1H, J=7 Hz, J=2Hz, CH—N), 6.14 (d, 1H, J=7 Hz, CH—O), 6.90 (dd, 1H, J=7.3 Hz, J=5 Hz,ArH), 7.02 (d, 1H, J=2 Hz, OCH═N), 7.60 (dd, 1H, J=7.3 Hz, J=1.7 Hz,ArH)), 8.13 (dd, 1H, J=5 Hz, J=1.8 Hz, ArH).

¹³C-NMR (CDCl₃, *): 24.3, 26.1, 46.3, 46.6, 53.5, 73.5, 78.1, 116.8,122.2, 135.2, 146.5, 155.3, 160.5 and 167.4.

MS-ESI m/z (% rel. Int.): 276.1 ([MH]⁺, 42).

HPLC: Method A, detection UV 254 nm, BAL 01014 RT=3.63 min, peak area97.2%.

trans-N N-Diethyl-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxamide SLA07194A

SLA 07194A was prepared in accordance with method D usingpyridine-4-carbaldehyde (1.14 mL, 9.52 mmol), KOH (0.54 g, 9.60 mmol) inmethanol (5 mL) and N,N-diethyl-2-isocyanoacetamide SLA 07184A (1.21 g,8.65 mmol). After work-up and column chromatography on florisil (ethylacetate)trans-N,N-diethyl-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxamide SLA07194A was obtained as a brown oil (0.25 g, 12% yield).

MW: 247.29; Yield: 12%; Brown Oil.

R_(f): 0.15 (AcOEt=100).

¹H-NMR (CDCl₃, *): 1.16-1.34 (m, 6H, CH₃), 3.30-3.80 (m, 4H, CH₂N), 4.60(dd, 1H, J=7.7 Hz, J=2.2 Hz, CH—N), 6.22 (d, 1H, J=7.7 Hz, CH—O), 7.06(d, J=2.2 Hz, CH═N), 7.23 (d, 2H, J=5.8 Hz, ArH), 8.61 (d, 2H, J=6.0 Hz,ArH).

Preparation of 2-chloropyridine-4-carbaldehyde SLA 07156 Methyl2-chloropyridine-4-carboxylate SLA 07150

2-Chloro-isonicotinic acid (5.10 g, 32.38 mmol) was dissolved inmethanol (150 mL). Thionyl chloride (12 mL) was added. This suspensionwas stirred 5 h at 70° C. and concentrated in vacuo. The residue wasdissolved in dichloromethane (250 mL) washed with a solution of 10%aqueous K₂CO₃ (2×150 mL) dried with MgSO₄, filtered and evaporated.Methyl 2-chloropyridine-4-carboxylate SLA 07150 was obtained as a yellowsolid (5.06 g, 91%).

MW: 171.58; Yield: 91%; Yellow Solid; Mp (° C.): 33.0.

R_(f): 0.80 (MeOH:CH₂Cl₂=10:90).

¹H-NMR (CDCl₃, *): 3.98 (s, 3H, CH₃), 7.78 (dd, 1H, J=5.1 Hz, J=1.3 Hz,ArH), 7.89 (d, 1H, J=0.6 Hz ArH), 8.55 (dd, 1H, J=5.1 Hz, J=0.6 Hz,ArH).

(2-Chloropyridin-4-yl)methanol SLA 07152

Methyl 2-chloropyridine-4-carboxylate (2.50 g, 14.60 mmol) was dissolvedin anhydrous THF (50 mL) and this solution was cooled to −78° C. underN₂ atmosphere. Diisobutylaluminium hydride 1.0 M in hexanes (63.3 mL,63.30 mmol) was added dropwise stabilizing the temperature between −50°C. and −70° C. The reaction mixture was stirred 1.5 h at −78° C. andallowed to stand at room temperature for 3 h. A solution of aqueous 10%NH₄Cl was slowly added and the mixture was extracted with ethyl acetate(3×300 mL). The combined organic layers were washed with water (3×20mL), brine (2×20 mL), dried over MgSO₄, filtered and evaporated.(2-Chloropyridin-4-yl)methanol SLA 07152 was obtained as a yellow oil(1.97 g, 94% yield).

MW: 143.71; Yield: 94%; Yellow Oil.

R_(f): 0.35 (EtOAc:cyclohexane=30:70).

¹H-NMR (CDCl₃, *): 2.95 (s broad, 1H, OH), 4.75 (s, 2H, CH₂O), 7.21 (dd,1H, J=5.1 Hz, J=1.2 Hz, ArH), 7.37 (d, 1H, J=1.2 Hz ArH), 8.29 (d, 1H,J=5.1 Hz, ArH).

MS-ESI m/z (rel. int.): 144.0 ([MH]⁺, 100).

HP LC: Method A, detection UV 254 nm, SLA 07152 RT=3.45 min, peak area99.9%.

2-Chloropyridine-4-carbaldehyde SLA 07156

In a 250 mL tricol equipped with a low temperature thermometer and twoequalizing dropping funnels was charged oxalyl dichloride (1.24 g, 9.81mmol) in dichloromethane (15 mL) and this solution was stirred under N₂at −78° C. The first equalizing dropping funnel was connected to anitrogen flow line and was charged with a solution of(2-chloropyridin-4-yl)methanol SLA 07152 (0.94 g, 6.54 mmol) withdichloromethane (15 mL). The other was charged with a solution ofdimethyl sulfoxide anhydrous (1.7 mL, 19.63 mmol) in dichloromethane (2mL) and this solution was added dropwise (25 min) in order to stabilizethe temperature between −60° C. and −70° C. At the end of the additionthe reaction solution was warmed to −60° C. over a period of 20 min thenthe solution of (2-chloropyridin-4-yl)methanol SLA 07152 was addeddropwise (50 min) keeping the temperature between −50° C. and −60° C. inthe reactor then the mixture reaction was warmed to −45° C. over aperiod of 30 min. The dropping funnel was washed with dichloromethane(2×5 mL) and charged with a solution of triethylamine (480 μl, 6.51mmol) in dichloromethane (4 mL) which was added (10 min) to the reactionmixture and finally the reaction flask was allowed to warm to 0° C. over10 min. The reaction solution was transferred to a 500 mL separatoryfunnel charged with 130 mL of a 5% aqueous NH₄Cl solution. The twophases were separated the aqueous phase was extracted withdichloromethane (3×50 mL) and the combined organic phases were washedwith 1 M aqueous phosphate buffer (pH=7; 4×100 mL), then dried overMgSO₄, filtered and evaporated. 2-Chloropyridine-4-carbaldehyde SLA07156 was obtained as an orange solid (0.740 g, 76% yield).

MW: 141.57; Yield: 76%; Orange Solid.

R_(f): 0.35 (EtOAc:cyclohexane=30:70).

¹H-NMR (CDCl₃, *): 7.65 (dd, 1H, J=5.0 Hz, J=1.3 Hz, ArH), 7.75 (d, 1H,J=1.3 Hz, ArH) 8.66 (d, 1H, J=5.0 Hz, ArH), 10.05 (s, 1H, CHO).

trans-(5-(2-Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07174

SLA 07174 was prepared in accordance with method D using2-chloropyridine-4-carbaldehyde SLA 07156 (0.12 g, 1.05 mmol), KOH (0.06g, 1.05 mmol) in methanol (10 mL) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.146 g, 1.05 mmol).The solution was stirred 24 h with continued cooling. After work-uptrans-(5-(2-chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07174 was obtained as a yellow solid (0.19 g, 66% yield).

MW: 279.72; Yield: 66%; Yellow Solid; Mp (° C.): 116.3.

¹H-NMR (CDCl₃, *): 1.86-2.07 (m, 4H, CH₂), 3.45-3.62 (m, 3H, CH₂N),3.93-4.01 (m, 1H, CH₂N), 4.50 (dd, J=8.0 Hz, J=2.3 Hz, 1H, CH—N), 6.19(d, 1H, J=8.0 Hz, CH—O), 7.02 (d, 1H, J=2.3 Hz, CH═N), 7.17 (td, 1H,J=5.1 Hz J=0.9 Hz, J=0.4 Hz, ArH), 7.29 (d, 1H, J=0.7 Hz, ArH), 8.38 (d,1H, J=5.1 Hz, ArH).

¹³C-NMR (CDCl₃, *): 22.5, 24.4, 44.9, 45.0, 74.3, 77.3, 117.2, 119.0,148.6, 150.4, 150.6, 153.1, 164.0.

trans-(5-(3-Bromopyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01028A

BAL 01028A was prepared in accordance with method D using3-bromo-4-pyridinecarboxaldehyde (1.010 g, 5.43 mmol), KOH (0.305 g,5.43 mmol) in methanol (5 mL) and 2-isocyano-1-pyrrolidin-1-yl-ethanoneBLE 04134 (0.75 g, 5.43 mmol). The mixture was stirred at 0° C. untilprecipitation and concentrated. The mixture was partitioned betweenEtOAc (50 ml) and H₂O (25 ml). The aqueous layer was extracted twicewith EtOAc (25 mL). The EtOAc fractions were combined, washed twice withbrine (2×25 mL), dried over MgSO₄ and filtered. After evaporation andtrans-(5-(3-bromopyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01028A was obtained (1.20 g, 68% yield) as a white solid.

MW: 324.17; Yield: 68%; White Solid; Mp (° C.): 160.8.

Rf: 0.25 (EtOAc=100).

¹H NMR (CDCl₃, *): 1.82-2.08 (m, 4H, 2×CH₂), 3.45-3.65 (m, 3H, CH₂N),3.80-3.92 (m, 1H, CH₂N), 4.60 (dd, 1H, J=2.1 Hz, J=6.1 Hz, CH—N), 6.30(d, 1H, J=6.1 Hz, CH—O), 7.10 (d, 1H, J=2.1 Hz, OCH═N), 7.30 (d, 1H,J=5.0 Hz, ArH)), 8.55 (d, 1H, J=5.0 Hz, ArH), 8.72 (s, 1H, ArH).

¹³C-NMR (CDCl₃, *): 24.3, 26.0, 46.4, 46.6, 74.5, 79.6, 118.6, 121.1,148.3, 148.8, 152.1, 155.1, 166.2.

MS-ESI m/z (% rel. Int.): 324.1/326.1 ([MH]⁺, 50/50), 239.0 (100).

HPLC: Method A, detection UV 254 nm, BAL 01028A RT=3.50 min, peak area96.8%.

trans-(5-(3-Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01028B

BAL 01028B was prepared in accordance with method D using2-isocyano-1-pyrrolidin-1-yl-ethanone BLE 04134 (0.75 g, 5.43 mmol), KOH(0.305 g, 5.43 mmol) in methanol (5 mL) and 3-chloro-isonicotinaldehyde(0.769 g, 5.43 mmol). The solution was stirred 3 h at 0° C.trans-(5-(3-Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01028B (1.20 g, 65% yield) was obtained as a white solid.

MW: 279.72; Yield: 65%; White Solid; Mp (° C.): 162.

Rf: 0.25 (EtOAc=100).

¹H NMR (CDCl₃, *): 1.82-2.08 (m, 4H, CH₂), 3.45-3.65 (m, 3H, CH₂N),3.82-3.93 (m, 1H, CH₂N), 4.62 (dd, 1H, J=2.1 Hz, J=6.1 Hz, CH—N), 6.38(d, 1H, J=6.1 Hz, CH—O), 7.08 (d, 1H, J=2.1 Hz, OCH═N), 7.33 (d, 1H,J=5.0 Hz, ArH), 8.52 (d, 1H, J=5.0 Hz, ArH), 8.59 (s, 1H, ArH).

¹³C-NMR (CD₃OD, *): 24.3, 26.0, 46.4, 46.6, 74.4, 77.9, 120.6, 128.8,146.6, 148.3, 149.7, 155.0, 166.1.

MS-ESI m/z (% rel. Int.): 280.1/282.1 ([MH]⁺, 39/14).

HPLC: Method A, detection UV 254 nm, BAL 01028B RT=3.47 min, peak area97.2%.

trans-(5-(2-Chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 07158

SLA 07158 was prepared in accordance with method D using2-chloropyridine-4-carbaldehyde SLA 07156 (0.47 g, 3.31 mmol), KOH(0.184 g, 3.33 mmol) in methanol (10 mL) and2-Isocyano-1-(2H-pyrrol-1(5H)-yl)ethanone SLA 07178 (0.410 g, 3.01mmol). The solution was stirred 2 h with continued cooling. Afterwork-up and column chromatography on florisil (EtOAc),trans-(5-(2-chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 07158 was obtained as a yellow solid (0.597 g, 84

MW: 277.71; Yield: 84%; Yellow Solid; Mp (° C.): 90.2.

R_(f): 0.10 (EtOAc).

¹H-NMR (CDCl₃, *): 4.26-4.37 (m, 3H, CH₂N), 4.48-4.52 (dd, 1H, J=2.3 Hz,J=8.0 Hz, CH—N), 4.75-4.85 (m, 1H, CH₂N), 5.80-5.95 (m, 2H, CH═CH), 6.20(d, 1H, J=8 Hz, CH—O), 7.02 (d, J=2.3 Hz, CH═N), 7.17 (td, 1H, J=5.1 Hz,J=0.8 Hz, J=0.6 Hz, ArH), 7.30 (t, 1H, J=0.6 Hz, ArH), 8.38 (d, 1H,J=5.1 Hz, ArH).

¹³C-NMR (CDCl₃, *): 53.4, 53.9, 75.8, 78.9, 118.9, 120.7, 125.2, 125.4,150.3, 151.9, 152.3, 154.

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 07180

SLA 07158 was prepared in accordance with method D usingpyridine-4-carbaldehyde (0.293 mL, 2.40 mmol), KOH (0.13 g, 2.32 mmol)in methanol (10 mL) and 2-isocyano-1-(2H-pyrrol-1(5H)-yl)ethanone SLA07178 (0.301 g, 2.20 mmol). The solution was stirred 2 h with continuedcooling. After work-up and column chromatography on florisil (EtOAc),trans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 07180 was obtained (0.284 g, 53% yield) as a yellow oil.

MW: 243.26; Yield: 53%; Yellow Oil.

R_(f): 0.15 (AcOEt).

¹H-NMR (CDCl₃, *): 4.28-4.33 (m, 3H, CH₂N), 4.52-4.56 (dd, 1H, J=7.8 Hz,J=2.2 Hz, CH—N), 4.73-4.82 (m, 1H, CH₂N), 5.80-5.93 (m, 2H, CH═CH), 6.18(d, 1H, J=7.8 Hz, CH—O), 7.08 (d, J=2.2 Hz, CH═N), 7.27 (d, 2H, J=6.0Hz, ArH), 8.59 (d, 2H, J=6.0 Hz, ArH).

¹³C-NMR (CDCl₃, *): 53.6, 53.9, 75.8, 79.7, 120.2, 125.3, 125.6, 148.7,150.5, 155.3, 166.1.

Preparation of Compound 20, Compound 21, Compound 22, Compound 23,Compound 24, Compound 25, Compound 26, Compound 27, Compound 28,Compound 29, Compound 30, Compound 31, Compound 32, Compound 34,Compound 35, Compound 36, Compound 37, Compound 38, Compound 39,Compound 40, Compound 41, Compound 42, Compound 43, Compound 44,Compound 45, Compound 46, Compound 48, Compound 49, and Compound 50.General method for oxazolines acidic hydrolysis: method E:

DL-threo-2-Amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 20

To a solution oftrans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110B (0.932 g, 3.80 mmol) in methanol (10 mL) was addedhydrochloric acid 37% (1.2 mL). After heating (50° C.) the mixture for2.25 h the reaction mixture was concentrated and the crude product wascoevaporated twice with ethyl acetate. After trituration with ethylacetate, filtration and dryingDL-threo-2-amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 20 was obtained as a white solid (1.10 g, 94%yield).

MW: 308.2; Yield: 94%; White Solid; Mp (° C.): 123.4.

¹H-NMR (CD₃OD, *): 1.65-2.00 (m, 4H, 2×CH₂), 2.82-3.11 (m, 1H, —CH₂N),3.30-3.57 (m, 2H, CH₂N), 3.57-3.77 (m, 1H, CH₂N), 4.54 (d, 1H, J=5.3 Hz,CH—N), 5.38 (d, 1H, J=5.3 Hz, CH—O), 8.15 (dd, 1H, J=7.6 Hz, J=5.0 Hz,ArH), 8.68 (d, 1H, J=7.6 Hz, ArH), 8.89 (d, 1H, J=7.6 Hz, ArH), 8.96 (s,1H, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 26.9, 47.7, 48.2, 58.1, 69.6, 128.7, 141.5,141.6, 143.1, 146.5, 165.4.

DL-threo-2-Amino-3-hydroxy-3-(pyridin-2-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 21

Compound 21 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04110 B (0.44 g, 1.79 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (10 mL). After 2.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-2-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 21 was obtained as a yellow solid (0.44 g, 84%yield).

MW: 308.28; Yield: 84%; Yellow Solid.

¹H-NMR (CD₃OD, *): 1.75-2.01 (m, 4H, 2×CH₂), 3.10-3.22 (m, 1H, CH₂N),3.39-3.60 (m, 2H, CH₂N), 3.63-3.75 (m, 1H, CH₂N), 4.71 (d, 1H, J=5.0 Hz,CH—N), 5.55 (d, 1H, J=5.0 Hz, CH—O), 8.05 (t, 1H, J=6.4 Hz, ArH), 8.13(d, 1H, J=8.0 Hz, ArH), 8.61 (t, 1H, J=8.0 Hz, ArH), 8.84 (d, 1H, J=5.6Hz, ArH).

DL-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22

Compound 22 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneCompound 19 (0.750 g, 3.07 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (10 mL). After 3.0 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 was obtained as a white solid (0.935 g, 99%yield).

MW: 308.28; Yield: 99%; White Solid; Mp (° C.): 117.0.

¹H-NMR (CD₃OD, *): 1.75-2.03 (m, 4H, 2×CH₂), 2.93-3.08 (m, 1H, CHN),3.32-3.75 (m, 3H, 2×CH₂), 4.54 (d, 1H, J=5.9 Hz, CH—N), 5.40 (d, 1H,J=5.9 Hz, CH—O), 8.21 (d, 2H, J=5.8 Hz, ArH), 8.94 (d, 2H, J=5.8 Hz,ArH).

MS-ESI m/z (% rel. int.): 236.1 ([MH]⁺, 17), 219 (25), 148 (100).

HPLC: Method A, detection UV 254 nm, Compound 22 RT=0.8 min, peak area96.3%.

DL-threo-2-Amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 23

Compound 23 was prepared following method E withtrans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124A (0.486 g, 1.94 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 23 was obtained as a white solid (0.480 g, 89.5%yield).

MW: 276.7; Yield: 89.5%; White Solid; Mp (° C.): 227.4.

¹H-NMR (CD₃OD, *): 1.47-1.88 (m, 4H, 2×CH₂), 2.31-2.46 (m, 1H, CH₂N),3.18-3.46 (m, 3H, CH₂N), 4.16 (d, 1H, J=9.0 Hz, CH—N), 4.97 (d, 1H,J=9.0 Hz, CH—O), 7.14 (dd, 1H, J=4.9 Hz, J=1.1 Hz, ArH), 7.40-7.50 (m,2H, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 26.7, 47.3, 47.6, 59.2, 70.6, 124.1, 127.1,127.7, 142.3, 166.3.

DL-threo-2-Amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-2-yl)propan-1-onehydrochloride Compound 24

Compound 24 was prepared following method E withtrans-(4,5-dihydro-5-(thiophen-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124B (0.677 g, 2.70 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-2-yl)propan-1-onehydrochloride Compound 24 was obtained as a white solid (0.630 g, 84.5%yield).

MW: 276.7; Yield: 84.5%; White Solid; Mp (° C.): 183.2.

¹H-NMR (CD₃OD, *): 1.49-1.90 (m, 4H, 2×CH₂), 2.36-2.48 (m, 1H, CH₂N),3.20-3.48 (m, 3H, CH₂N), 4.18 (d, 1H, J=9.1 Hz, CH—N), 5.14 (d, 1H,J=9.1 Hz, CH—O), 7.00-7.08 (m, 2H, ArH), 7.45 (dd, 1H, J=4.9 Hz, J=1.6Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 26.8, 47.3, 47.7, 59.6, 70.5, 126.3, 127.0,128.2, 144.5, 166.1.

DL-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiazol-2-yl)propan-1-onedihydrochloride Compound 25

Compound 25 was prepared following method E withtrans-(4,5-dihydro-5-(thiazol-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124C (0.558 g, 2.22 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiazol-2-yl)propan-1-onedihydrochloride Compound 25 was obtained as a pale yellow solid (0.532g, 76.5% yield).

MW: 276.7; Yield: 76.5%; Pale Yellow Solid; Mp (° C.): 145.8.

¹H-NMR (CD₃OD, *): 1.75-2.00 (m, 4H, 2×CH₂), 3.05-3.17 (m, 1H, —CH₂N),3.36-3.58 (m, 2H, CH₂N), 3.58-3.70 (m, 1H, CH₂N), 4.67 (d, 1H, J=5.4 Hz,CH—N), 5.49 (d, 1H, J=5.4 Hz, CH—O), 7.84 (d, 1H, J=3.4 Hz ArH), 7.99(d, 1H, J=3.4 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 27.0, 47.7, 48.0, 57.5, 69.9, 123.6, 142.1,165.3, 173.3.

DL-threo-2-Amino-3-(3a,7a-dihydrobenzo[b]thiophen-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 26

Compound 26 was prepared following method E withtrans-(5-(benzo[b]thiophen-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04124D (1.050 g, 3.49 mmol), hydrochloric acid 37% (1.2 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-(3a,7a-dihydrobenzo[b]thiophen-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 26 was obtained as a white solid (0.970 g, 85%yield).

MW: 326.84; Yield: 85%; White Solid; Mp (° C.): 207.0.

¹H-NMR (CD₃OD, *): 0.92-1.09 (m, 2H, 2×CH₂), 1.42-1.60 (m, 2H, 2×CH₂),1.83-1.98 (m, 1H, CH₂N), 2.76-2.91 (m, 1H, CH₂N), 3.06-3.25 (m, 2H,—CH₂N), 4.30 (d, 1H, J=9.5 Hz, CH—N), 5.29 (d, 1H, J=9.5 Hz, CH—O),7.35-7.43 (m, 2H, ArH), 7.78-7.89 (m, 2H, ArH), 7.90-7.97 (m, 1H, ArH).

¹³C-NMR (CD₃OD, *): 24.5, 26.4, 47.3, 47.4, 59.0, 69.5, 123.1, 124.0,125.4, 126.1, 126.8, 136.6, 138.3, 141.9, 166.1.

DL-threo-2-Amino-3-(furan-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 27

Compound 27 was prepared following method E withtrans-(5-(furan-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130B (0.800 g, 3.41 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-(furan-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 27 was obtained as a white solid (0.738 g, 83%yield).

MW: 260.72; Yield: 83%; White Solid; Mp (° C.): 218.0.

¹H-NMR (CD₃OD, *): 1.62-1.95 (m, 4H, 2×CH₂), 2.82-2.95 (m, 1H, CH₂N),3.22-3.38 (m, 1H, CH₂N), 3.39-3.55 (m, 2H, CH₂N), 4.19 (d, 1H, J=8.4 Hz,CH—N), 4.90 (d, 1H, J=8.4 Hz, CH—O), 6.49 (m, 1H, ArH), 7.52-7.57 (m,2H, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 26.7, 47.4, 48.0, 58.7, 67.2, 109.8, 125.9,142.0, 145.2, 166.3.

DL-threo-2-Amino-3-hydroxy-3-(naphthalen-2-yl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 28

Compound 28 was prepared following method E withtrans-(4,5-dihydro-5-(naphthalen-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130C (0.745 g, 2.53 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(naphthalen-2-yl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 28 was obtained as a white solid (0.706 g, 87%yield).

MW: 320.81; Yield: 87%; White Solid; Mp (° C.): 173.8.

¹H-NMR (CD₃OD, *): 0.93-1.10 (m, 1H, CH₂), 1.20-1.37 (m, 1H, CH₂),1.44-1.71 (m, 2H, CH₂), 1.99-2.10 (m, 1H, CH₂N), 3.11-3.26 (m, 2H,CH₂N), 3.31-3.41 (m, 1H, CH₂N), 4.23 (d, 1H, J=9.1 Hz, CH—N), 5.06 (d,1H, J=9.1 Hz, CH—O), 7.50-7.63 (m, 3H, ArH), 7.87-7.97 (m, 4H, ArH).

¹³C-NMR (CD₃OD, *): 24.6, 26.3, 47.2, 47.5, 59.4, 74.3, 125.1, 126.9,127.7, 127.8, 128.8, 129.0, 129.4, 134.5, 135.0, 138.0, 166.4.

DL-threo-2-Amino-3-hydroxy-3-(naphthalen-1-yl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 29

Compound 29 was prepared following method E withtrans-(4,5-dihydro-5-(naphthalen-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04130D (0.794 g, 2.69 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(naphthalen-1-yl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 29 was obtained as a white solid (0.768 g, 89%yield).

MW: 320.81; Yield: 89%; White Solid; Mp (° C.): 177.8.

¹H-NMR (CD₃OD, *): 0.71-0.91 (m, 2H, CH₂), 1.29-1.51 (m, 3H, CH₂),2.54-2.67 (m, 1H, CH₂N), 2.88-3.02 (m, 1H, CH₂N), 3.02-3.16 (m, 1H,CH₂N), 4.27 (d, 1H, J=9.8 Hz, CH—N), 5.67 (d, 1H, J=9.8 Hz, CH—O),7.50-7.61 (m, 3H, ArH), 7.90-7.98 (m, 3H, ArH), 8.08-8.14 (m, 1H, ArH).

¹³C-NMR (CD₃OD, *): 24.4, 26.2, 47.1, 47.3, 59.5, 70.3, 124.0, 126.5(2×C), 127.2, 127.4, 129.9, 130.4, 132.1, 135.0, 137.1, 166.1.

DL-threo-2-Amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(quinolin-2-yl)propan-1-onedihydrochloride Compound 30

Compound 30 was prepared following method E withtrans-(4,5-dihydro-5-(quinolin-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136B (0.923 g, 3.13 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (15 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(quinolin-2-yl)propan-1-onedihydrochloride Compound 30 was obtained as a yellow solid (1.098 g, 98%yield).

MW: 358.26; Yield: 98%; Yellow Solid; Mp (° C.): 131.5.

¹H-NMR (CD₃OD, *): 1.69-2.07 (m, 4H, CH₂), 3.16-3.34 (m, 3H, CH₂),3.37-3.60 (m, 2H, CH₂N), 3.77-3.88 (m, 1H, CH₂—N), 5.85 (d, 1H, J=4.9Hz, CH—O), 8.03 (t, 1H, J=7.6 Hz, ArH), 8.17-8.30 (m, 2H, ArH), 8.40 (d,1H, J=8.3 Hz, ArH), 8.56 (d, 1H, J=8.6 Hz, ArH), 9.25 (d, 1H, J=8.6 Hz,ArH), not seen under H₂O (d, 1H, CH—NH₂).

¹³C-NMR (CD₃OD, *): 24.9, 27.0, 47.9, 48.2, 57.3, 70.3, 121.5, 122.5,130.4, 130.5, 131.5, 136.5, 140.2, 148.5, 157.8, 164.8.

DL-threo-2-Amino-3-hydroxy-3-(isoquinolin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 31

Compound 31 was prepared following method E withtrans-(4,5-dihydro-5-(isoquinolin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136C (0.597 g, 2.02 mmol), hydrochloric acid 37% (0.4 mL) andmethanol (10 mL). After 3.5 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(isoquinolin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 31 was obtained as a off white solid (0.716 g,99% yield).

MW: 358.27; Yield: 99%; Off White Solid; Mp (° C.): 158.5.

¹H-NMR (CD₃OD, *): 1.04-1.32 (m, 2H, CH₂), 1.51-1.72 (m, 2H, CH₂),2.05-2.20 (m, 1H, CH₂N), 2.68-2.80 (m, 1H, CH₂N), 3.20-2.47 (m, 1H,CH₂N), 4.57 (d, 1H, J=8.5 Hz, CH—NH₂), 5.99 (d, 1H, J=8.5 Hz, CH—OH),8.09 (t, 1H, J=8.6 Hz, ArH), 8.27 (t, 1H, J=8.6 Hz, ArH), 8.38 (d, 1H,J=8.6 Hz, ArH), 8.45 (d, 1H, J=8.6 Hz, ArH), 8.55 (d, 1H, J=5.6 Hz,ArH), 9.35 (d, 1H, J=5.6 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.6, 26.5, 47.5, 48.0, 58.2, 69.1, 122.3, 122.6,125.9, 127.7, 131.7, 136.7, 138.9, 146.1, 159.4, 165.2.

N-(DL-threo-1-hydroxy-3-oxo-3-(pyrrolidin-1-yl)-1-(quinolin-3-yl)propan-2-yl)formamidehydrochloride Compound 32

Compound 32 was prepared following method E withtrans-(4,5-dihydro-5-(quinolin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01016 (0.905 g, 3.41 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 2 h at RT and work-upN-(DL-threo-1-hydroxy-3-oxo-3-(pyrrolidin-1-yl)-1-(quinolin-3-yl)propan-2-yl)formamidehydrochloride Compound 32 was obtained as a white solid (240 mg, 20.0%yield).

MW: 349.81; Yield: 20.0%; White Solid; Mp (° C.): 203.2.

¹H NMR (CD₃OD, *): 1.78-2.09 (m, 4H, CH₂), 3.35-3.58 (m, 2H, CH₂N),3.58-3.80 (m, 2H, CH₂N), 5.28 (d, 1H, J=4 Hz, CH—N), 5.51 (d, 1H, J=4Hz, CH—O), 8.00 (t, 2H, J=7.1 Hz, ArH)), 8.18 (t, 1H, J=6.9 Hz, ArH),8.26 (d, 1H, J=8.6 Hz, ArH), 8.36 (d, 1H, J=8.3 Hz, ArH), 9.18 (s, 1H,CHO), 9.26 (s, 1H, ArH).

¹³C-NMR (CD₃OD, *): 25.1, 27.0, 47.5, 48.3, 55.5, 71.2, 121.4, 129.9,130.6, 131.5, 136.2, 137.3, 138.5, 145.3, 145.8, 163.4, 168.7.

MS-ESI m/z (% rel. Int.): 314 ([MH]⁺, 50), 158.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 32 RT=3.36 min, peak area99.9%.

DL-threo-2-Amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(quinolin-3-yl)propan-1-onedihydrochloride Compound 33

Compound 33 was prepared following method E withtrans-(4,5-dihydro-5-(quinolin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01016 (0.91 g, 3.41 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (10 mL). After 3 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(quinolin-3-yl)propan-1-onedihydrochloride Compound 33 (678 mg, 55% yield) was obtained as a whitesolid.

MW: 358.26; Yield: 55%; White Solid; Mp (° C.): 190.9.

¹H NMR (CD₃OD, *): 1.57-1.80 (m, 2H, CH₂), 1.80-1.99 (m, 2H, CH₂),3.01-3.20 (m, 1H, CH₂N), 3.35-3.61 (m, 2H, CH₂N), 3.61-3.82 (m, 1H,CH₂N), 4.70 (d, 1H, J=5.0 Hz, CH—N), 5.58 (d, 1H, J=5.0 Hz, CH—O),7.96-8.11 (m, 1H, ArH), 8.18-8.29 (m, 1H, ArH), 8.29-8.38 (m, 1H, ArH),8.38-8.49 (m, 1H, ArH), 9.28 (s, 1H, ArH), 9.34 (s, 1H, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 26.9, 47.8, 48.3, 58.2, 69.8, 121.8, 130.0,130.8, 131.9, 135.4, 136.9, 139.3, 145.1, 146.2, 165.6.

MS-ESI m/z (% rel. Int.): 286.2 ([MH]⁺, 100).

HPLC: Method A, detection UV 254 nm, Compound 33 RT=3.15 min, peak area97.0%.

DL-threo-2-Amino-3-(2-chloropyridin-4-yl)-3-hydroxy-1-(2H-pyrrol-1(5H)-yl)propan-1-onedihydrochloride Compound 34

Compound 34 was prepared following method E withtrans-(5-(2-chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 07158 (0.597 g, 2.02 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (10 mL). After 2 h at room temperature and work-upDL-threo-2-amino-3-(2-chloropyridin-4-yl)-3-hydroxy-1-(2H-pyrrol-1(5H)-yl)propan-1-onedihydrochloride Compound 34 (0.656 mg, 91% yield) was obtained as a paleyellow solid.

MW: 340.63; Yield: 91%; Pale Yellow Solid; Mp (° C.): 196.2.

¹H-NMR (CD₃OD, *): 3.45-3.50 (m, 1H, CH₂N), 4.04-4.15 (m, 1H, CH₂N),4.22-4.36 (m, 3H, CH₂N & CHNH₂), 5.05 (d, 1H, J=7.1 Hz, —CHO), 5.71 (d,1H, J=4.3 Hz, CH═CH), 5.84 (d, 1H, J=4.3 Hz, CH═CH), 7.47 (d, 1H, J=5.0Hz, ArH), 7.57 (s, 1H, ArH), 8.39 (d, 1H, J=5.0 Hz, ArH).

¹³C-NMR (CD₃OD, *): 54.3, 54.5, 58.2, 71.7, 122.0, 123.5, 125.7, 126.3,151.0, 152.8, 154.0, 165.8.

DL-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(piperidin-1-yl)propan-1-onedihydrochloride Compound 35

Compound 35 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(piperidin-1-yl)methanoneSLA 07122A (0.33 g, 1.27 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (10 mL). After 3 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(piperidin-1-yl)propan-1-onedihydrochloride Compound 35 was obtained as a yellow solid (0.375 g, 91%yield).

MW: 322.31; Yield: 91%; Yellow Solid; Mp (° C.): 145.

¹H-NMR (CD₃OD, *): 1.05-1.17 (m, 1H, CH₂) 1.28-1.65 (m, 5H, CH₂),2.75-3.00 (m, 1H, CH₂N), 3.10-3.22 (m, 1H, CH₂N), 3.23-3.38 (m, 1H,CH₂N), 3.53-3.65 (m, 1H, CH₂N), 4.68 (d, 1H, J=5.8 Hz, CHNH₂), 5.14 (d,1H, J=5.8 Hz, CHO), 8.06 (d, 2H, J=6.0 Hz, ArH), 8.79 (d, 2H, J=6.5 Hz,ArH).

DL-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(morpholin-1-yl)propan-1-onedihydrochloride Compound 36

Compound 36 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(morpholino)methanone SLA07124A (0.146 g, 0.56 mmol), hydrochloric acid 37% (1.0 mL) and methanol(10 mL). After 3 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(morpholin-1-yl)propan-1-onedihydrochloride Compound 36 was obtained as a pale yellow solid (0.143g, 89% yield).

MW: 287.78; Yield: 89%; Pale Yellow Solid; Mp (° C.): 115.9.

¹H-NMR (CD₃OD, *): 3.32-3.82 (m, 8H, 4×CH₂), 5.41 (d, 1H, J=5.0 Hz,CHO—), 8.28 (d, 2H, J=5.9 Hz, ArH), 8.97 (d, 2H, J=5.8 Hz, ArH), CHNH₂not seen.

DL-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(piperazin-1-yl)propan-1-onetrihydrochloride Compound 37

Compound 37 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-tert-butyloxycarbonyl-piperazin-1-yl)methanoneSLA 07124B (0.31 g, 0.86 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (10 mL). After 3 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(piperazin-1-yl)propan-1-onetrihydrochloride Compound 37 was obtained as a yellow solid (0.303 g,71%).

MW: 359.8; Yield: 71%; Yellow Solid; Mp (° C.): 201.4.

R_(f): 0.20 (CH₂Cl₂:MeOH=90:10), free base.

¹H-NMR (CD₃OD, *): 3.31-3.48 (m, 4H, 2×CH₂), 3.63-3.90 (m, 2H, CH₂N),4.00-4.35 (m, 2H, CH₂N), 5.15 (d, 1H, J=4.5 Hz, CHNH₂), 5.58 (d, 1H,J=4.5 Hz, CHO), 8.38 (d, 2H, J=6.4 Hz, ArH), 9.04 (d, 2H, J=6.5 Hz,ArH).

DL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-thiomorpholinopropan-1-onedihydrochloride Compound 38

Compound 38 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(thiomorpholino)methanoneSLA 07132 (0.926 g, 3.36 mmol), hydrochloric acid 37% (1.1 mL) andmethanol (10 mL). After 3 h at 50° C. and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-thiomorpholinopropan-1-onedihydrochloride Compound 38 was obtained as a pale yellow solid (1.1 g,99% yield).

MW: 340.35; Yield: 99%; Pale Yellow Solid; Mp (° C.): 200.6.

¹H-NMR (CD₃OD, *): 2.42-2.52 (m, 1H, CH₂), 2.53-2.70 (m, 1H, CH₂),2.70-2.90 (m, 2H, CH₂), 3.45-3.71 (m, 2H, CH₂N), 3.87-4.00 (m, 1H,CH₂N), 4.18-4.28 (m, 1H, CH₂N), 5.44 (d, 1H, J=5.1 Hz, CHO), 8.34 (d,2H, J=5.9 Hz, ArH), 9.03 (d, 2H, J=5.6 Hz, ArH), —CHNH₂ not seen (underH₂O).

¹³C-NMR (CD₃OD, *): 27.9, 28.7, 46.6, 50.0, 56.0, 71.3, 126.9 (2×C),143.2 (2×C), 161.3, 165.7.

DL-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(2H-pyrrol-1(5H)-yl)propan-1-onedihydrochloride Compound 39

Compound 39 was prepared following method E withtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 07180 (0.276 g, 1.14 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (10 mL). After 2.5 h at RT and work-upDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(2H-pyrrol-1(5H)-yl)propan-1-onedihydrochloride Compound 39 was obtained (343 mg, 99% yield) as a whitesolid.

MW: 306.27; Yield: 99%; White Solid; Mp (° C.): 186.3.

¹H-NMR (CD₃OD, *): 3.91-4.02 (m, 1H, CH—NH₂), 4.09-4.21 (m, 1H, CH₂),4.27-4.41 (m, 1H, CH₂), 4.44-4.59 (m, 1H, CH₂), 4.55 (d, 1H, J=5.7 Hz,CH₂N), 5.46 (d, 1H, J=5.7 Hz, CHO), 5.80-5.90 (m, 2H, CH═CH), 8.24 (d,1H, J=6.3 Hz, ArH), 8.93 (d, 1H, J=5.7 Hz, ArH).

¹³C-NMR (CD₃OD, *): 54.6, 54.7, 57.6, 71.0, 125.9, 126.4, 126.8 (2×C),143.1 (2×C), 161.6, 165.5.

MS-ESI m/z (% rel. Int.): 234.1 ([MH]⁺, 5), 137.1 (100).

DL-threo-2-Amino-N,N-diethyl-3-hydroxy-3-(pyridin-4-yl)propanamidedihydrochloride Compound 40

Compound 40 was prepared following method E withtrans-N,N-diethyl-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxamidediethylamide SLA 07194A (254 mg, 1.03 mmol), hydrochloric acid 37% (1.0mL) and methanol (10 mL). After 2 h at RT and work-upDL-threo-2-amino-N,N-diethyl-3-hydroxy-3-(pyridin-4-yl)propanamidedihydrochloride Compound 40 was obtained (212 mg, 67% yield) as a paleyellow solid.

MW: 310.30; Yield: 67%; Pale Yellow Solid; Mp (° C.): 159.6° C.

R_(f): 0.10 (CH₂Cl₂:MeOH=90:10), free base.

¹H-NMR (CD₃OD, *): 1.01-1.12 (m, 6H, 2×CH₃), 3.01-3.31 (m, 3H, CH₂),3.40-3.52 (m, 1H, CH₂), 4.64 (d, 1H, J=6.8 Hz, CHN), 5.31 (d, 1H, J=6.8Hz, CHO), 8.22 (d, 1H, J=6.4 Hz, ArH), 8.94 (d, 1H, J=6.4 Hz, ArH).

¹³C-NMR (CD₃OD, *): 12.9, 14.4, 42.1, 43.4, 55.9, 72.1, 126.9 (2×C),143.3 (2×C), 161.5, 166.1.

MS-ESI m/z (% rel. Int.): 238.1 ([MH]⁺, 5), 137.1 (100).DL-threo-2-Amino-3-(2-chloropyridin-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 41.

Compound 41 was prepared following method E withtrans-(5-(2-chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 07174 (0.179 g, 0.64 mmol), hydrochloric acid 37% (1.0 mL) andmethanol (7 mL). After 2 h at RT and work-upDL-threo-2-amino-3-(2-chloropyridin-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 41 was obtained (212 mg, 67 (142 mg, 65% yield)as a pale yellow solid.

MW: 342.65; Yield: 65%; Pale Yellow Solid; Mp (° C.): 184.3.

R_(f): 0.15 (CH₂Cl₂:MeOH=90:10), free base.

¹H-NMR (CD₃OD, *): 1.50-1.90 (m, 4H, 2×CH₂), 2.50-2.61 (m, 1H, CH₂N),3.25-3.38 (m, 1H, CH₂N), 3.40-3.53 (m, 2H, CH₂N), 4.26 (d, 1H, J=7.7 Hz,CHN), 4.99 (d, 1H, J=7.7 Hz, CHO), 7.45 (d, 1H, J=4.5 Hz, ArH), 7.53 (s,1H, ArH), 8.41 (d, 1H, J=4.9 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 26.7, 47.5, 48.0, 58.4, 71.9, 122.0, 123.5,151.1, 152.7, 154.0, 165.7.

MS-ESI m/z (% rel. Int.): 270.1/272.1 ([MH]⁺, 40/13), 171.0/172.0(100/32).

DL-threo-2-Amino-3-(3-bromopyridin-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 42

Compound 42 was prepared following method E withtrans-(5-(3-bromopyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01028A (1.141 g, 3.52 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (15 mL). After 3 h at 50° C. and work-upDL-threo-2-amino-3-(3-bromopyridin-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 42 was obtained as a white solid (667 mg, 49%yield).

MW: 387.10; Yield: 49.0%; White Solid; Mp (° C.): 216.3.

¹H NMR (CD₃OD, *): 1.73-1.99 (m, 4H, 2×CH₂), 3.01-3.05 (m, 1H, CH₂N),3.44-3.51 (m, 2H, CH₂N), 3.60-3.73 (m, 1H, CH₂N), 4.60 (d, 1H, J=5.5 Hz,CH—N), 5.54 (d, 1H, J=5.5 Hz, CH—O), 8.26 (d, 1H, J=5.7 Hz, ArH)), 8.86(d, 1H, J=5.7 Hz, ArH), 9.10 (s, 1H, ArH).

¹³C-NMR (CD₃OD, *): 24.8, 27.1, 56.4, 70.1, 122.7, 127.6, 145.1, 148.4,156.8, 165.0, 2×C not seen.

MS-ESI m/z (% rel. Int.): 314.1/316.1 ([MH]⁺, 35/35), 215.0/217 (50/50).

HPLC: Method A, detection UV 254 nm, Compound 42 RT=3.08 min, peak area92.8%.

DL-threo-2-Amino-3-(3-chloropyridin-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 43

Compound 43 was prepared following method E withtrans-(5-(3-chloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01028B (0.925 g, 3.31 mmol), hydrochloric acid 37% (0.6 mL) andmethanol (15 mL). After 2 h at 50° C. and work-upDL-threo-2-amino-3-(3-chloropyridin-4-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 43 was obtained as a white solid (599 mg, 53%yield).

MW: 342.65; Yield: 53%; White Solid; Mp (° C.): 214.0.

¹H NMR (CD₃OD, *): 1.75-2.02 (m, 4H, 2×CH₂), 3.13-3.25 (m, 1H, CH₂N),3.39-3.62 (m, 2H, CH₂N), 3.65-3.80 (m, 1H, CH₂N), 4.66 (d, 1H, J=4.5 Hz,CH—N), 5.66 (d, 1H, J=4.6 Hz, CH—O), 8.40 (d, 1H, J=5.8 Hz, ArH), 8.93(d, 1H, J=5.8 Hz, ArH), 9.13 (s, 1H, ArH).

¹³C-NMR (CD₃OD, *): 23.3, 25.6, 46.6, 46.8, 54.6, 66.5, 126.5, 132.4,141.7, 142.9, 155.6, 163.4.

MS-ESI m/z (% rel. Int.): 270/272 ([MH]⁺, 33/11), 171/173 (100/32).

HPLC: Method A, detection UV 254 nm, Compound 43 RT=2.80 min, peak area97.2%.

DL-threo-3-Hydroxy-1-oxo-3-(1-oxy-pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateBAL 01060

To a solution ofDL-threo-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamate(300 mg, 0.81 mmol, free base obtained from Compound 58 by K₂CO₃, CH₂Cl₂treatment) in dichloromethane (40 mL) was added MCPBA (350 mg, 2.03mmol). The resulting mixture was stirred overnight at room temperature.The mixture was concentrated and the crude product was purified bycolumn chromatography (EtOAc:MeOH=70:30).DL-threo-3-Hydroxy-1-oxo-3-(1-oxy-pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateBAL 01060 was obtained as a white solid (292 mg, 94% yield).

MW: 385.41; Yield: 94%; White Solid.

¹H NMR CDCl₃, *): 1.71-2.00 (m, 4H, 2×CH₂), 3.35-3.53 (m, 3H, CH₂N),3.54-3.68 (m, 1H, CH₂N), 4.65 (dd, 1H, J=9.6 Hz, J=2.0 Hz, CH—N),4.90-5.12 (m, 3H, CH₂O & OH), 5.20 (d, 1H, J=1.9 Hz, CH—O), 5.83 (d, 1H,J=9.6 Hz, NH), 7.20-7.40 (m, 7H, ArH), 8.08 (d, 2H, J=7.1 Hz, ArH).

DL-threo-2-Amino-3-hydroxy-3-(1-oxy-pyridin-4-yl)-1-pyrrolidin-1-yl-propan-1-onehydrochloride Compound 44

[2-Hydroxy-2-(1-oxy-pyridin-4-yl)-1-(pyrrolidine-1-carbonyl)-ethyl]-carbamicacid benzyl ester BAL 01060 (0.26 g, 0.67 mmol) was dissolved in a 6 Nhydrochloric acid solution (10 mL). The solution was stirred for 0.75 hat 100° C. The residue was concentrated, dissolved in MeOH:EtOAc=50:50and heated at reflux. After cooling, the mixture was evaporated,triturated in MeOH and filtered to obtainDL-threo-2-amino-3-hydroxy-3-(1-oxy-pyridin-4-yl)-1-pyrrolidin-1-yl-propan-1-onehydrochloride Compound 44 (65 mg, 33% yield) as a white solid.

MW: 287.74; Yield: 33%; White Solid; Mp (° C.): 178.5.

¹H NMR (D₂O, *): 1.55-1.93 (m, 4H, 2×CH₂), 2.65-3.80 (m, 1H, CH₂N),3.22-3.56 (m, 3H, CH₂N), 4.43 (d, 1H, J=7.6 Hz, CH—N), 5.19 (d, 1H,J=7.6 Hz, CH—O), 7.69 (d, 2H, J=6.1 Hz, ArH), 8.39 (d, 2H, J=6.9 Hz,ArH).

¹³C-NMR (D₂O, *): 24.1, 25.8, 47.3, 48.0, 57.4, 70.5, 125.7 (2×C), 139.9(2×C), 143.7, 165.1.

MS-ESI m/z (% rel. Int.): 252.1 ([MH]⁺, 18), 120.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 44 RT=0.8 min, peak area99.9%.

DL-threo-2-(Dimethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 45

DL-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (0.50 g, 1.62 mmol) and para formaldehyde(0.245 g, 8.11 mmol) were stirred in methanol (25 mL) for 10 min. Sodiumcyanoborohydride (0.612 g, 9.73 mmol) was added. The solution wasstirred 19 h at 50° C. and then concentrated. The residue waspartitioned between dichloromethane and water. The aqueous layer wasbasified with 1N sodium hydroxyde (pH=10). The organic layer wascombined with additional dichloromethane extracts, washed with aqueoussodium chloride and dried with MgSO₄. The crude product was purified bycolumn chromatography on silica (CH₂Cl₂:MeOH=95:05).DL-threo-2-(dimethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneSLA 07140 was obtained (187 mg, 44%) as a yellow oil. To a stirredsolution ofDL-threo-2-(dimethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneSLA 07140 (0.142 g, 0.54 mmol) in ethyl acetate (5 mL) was addeddropwise via syringe 4 mL of a solution of HCl in Et₂O (0.3 M). Thereaction mixture was stirred at 0° C. for 0.5 h. The precipitate wasfiltered, washed with Et₂O and dried.DL-threo-2-(Dimethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 45 was obtained (0.077 g, 43% yield) as a whitesolid.

MW: 336.34; Yield: 43%; White Solid; Mp (° C.): 201.0.

¹H-NMR (CD₃OD, *: 1.48-1.64 (m, 2H, CH₂), 1.65-1.83 (m, 2H, CH₂),2.60-2.72 (m, 1H, CH₂N), 3.15-3.33 (m, 1H, CH₂), 3.30-3.52 (m, 2H, CH₂),4.60 (d, 1H, J=8.4 Hz, CHNH₂), 5.41 (d, 1H, J=8.4 Hz, CHO), 8.10 (d, 2H,J=6.6 Hz, ArH), 8.84 (d, 2H, J=6.7 Hz, ArH).

DL-threo-2-Amino-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 46

To a stirred suspension ofDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (0.86 g, 2.80 mmol) in tetrahydrofuran (108mL) under nitrogen atmosphere was slowly added, in two portions, lithiumaluminium hydride (0.64 g, 16.82 mmol) at 0° C. The mixture reaction wasstirred at RT for 20 h and quenched by a slow, dropwise addition of 2 Naqueous sodium hydroxyde (8.4 mL, 6 eq). The yellow precipitate wasfiltered. The organic layer was washed by water (80 mL) and the organiclayer was removed and combined with additional ethyl acetate extracts(4×200 mL) and dried over MgSO₄, filtered and evaporated. The crudeproduct was purified by column chromatography on silica(CH₂Cl₂:MeOH:NH₃=94:05:01). After evaporation and dryingDL-threo-2-amino-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-1-olCompound 46 was obtained (0.075 g, 12% yield) as a pale yellow solid.

MW: 221.30; Yield: 12%; Pale Yellow Solid.

R_(f): 0.35 (CH₂Cl₂:MeOH:NH₃=90:08:02).

¹H-NMR (CD₃OD, *): 1.60-1.80 (m, 4H, 2×CH₂), 2.30-2.80 (m, 6H, 3×CH₂N),3.14-3.19 (m, 1H, CHNH₂), 4.68 (d, 1H, J=3.0 Hz, CHO), 7.30 (d, 2H,J=6.0 Hz, ArH), 8.55 (d, 2H, J=6.0 Hz, ArH).

¹³C-NMR (CD₃OD, *): 23.5 (2×C), 54.1, 54.7 (2×C), 60.1, 74.5, 121.4(2×C), 149.5 (2×C), 152.1.

MS-ESI m/z (rel. int.): 222.1 ([MH]⁺, 100), 205.0 (80), 189.0 (45),151.0 (70), 134.0 (42), 121.9 (100), 107.9 (40).

DL-threo-2-Amino-3-hydroxy-3-(2-methoxypyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 48

Trans-(4,5-Dihydro-5-(2-methoxypyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01014 (0.465 g, 1.69 mmol) was dissolved in methanol (6 mL). Thesolution of hydrochloric acid (37%, 0.3 mL) was added via a syringe atRT. The mixture was stirred for 3 h at RT. The residue was concentrated,dissolved in the minimum of MeOH, precipitated with EtOAc and filteredto obtain a white solidDL-threo-2-amino-3-hydroxy-3-(2-methoxypyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 48 (103 mg, 18.0% yield).

MW: 338.23; Yield: 18.0%; White Solid; Mp (° C.): 171.5.

¹H NMR (CD₃OD, *): 1.85-2.10 (m, 4H, CH₂), 3.30-3.82 (m, 4H, CH₂N), 4.26(s, 3H, OCH₃), 4.60 (d, 1H, J=3.7 Hz, CH—N), 5.45 (d, 1H, J=3.7 Hz,CH—O), 7.39 (dd, 1H, J=5.6 Hz, J=7.3, ArH), 8.32 (dd, 1H, J=5.6 Hz,J=7.3 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.9, 27.1, 47.8, 47.9, 56.4, 56.7, 66.0, 119.2,125.6, 141.5, 143.9, 160.6, 166.1.

MS-ESI m/z (% rel. Int.): 266.2 ([MH]⁺, 30), 248.2.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 48 RT=3.31 min, peak area97.9%.

3-(DL-threo-2-Amino-1-hydroxy-3-oxo-3-pyrrolidin-1-yl-propyl)-1H-pyridin-2-onehydrochloride Compound 49

Trans-(4,5-Dihydro-5-(2-methoxypyridin-3-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneBAL 01014 (0.684 g, 2.487 mmol) was dissolved in methanol (10 mL). Asolution of hydrochloric acid (37%, 0.6 mL) was added via syringe at RT.The mixture was stirred for 22 h at reflux. The residue wasconcentrated, triturated with EtOAc and filtered to obtain a yellow palesolid3-(DL-threo-2-amino-1-hydroxy-3-oxo-3-pyrrolidin-1-yl-propyl)-1H-pyridin-2-onehydrochloride Compound 49 (136 mg, 19.0% yield).

MW: 287.74; Yield: 19.0%; Yellow Pale Solid; Mp (° C.): 180.

¹H NMR (CD₃OD, *): 1.82-2.09 (m, 4H, 2×CH₂), 3.35-3.80 (m, 4H, 2×CH₂N),4.63 (s, 1H, CH—N), 5.17 (s, 1H, CH—O), 6.56 (t, 1H, ArH)), 7.5 (d, 1H,J=6.1 Hz, ArH), 7.86 (d, 1H, J=6.5 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.2, 26.0, 46.6, 46.6, 75.8, 79.7, 127.3, 127.5,127.9, 129.4, 130.0, 132.3, 133.2, 148.1, 148.4, 155.3, 166.2.

MS-ESI m/z (% rel. Int.): 252.1 ([MH]⁺, 18), 163.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 49 RT=1.13 min, peak area84.0%.

Preparation of Compound 51, Compound 52, Compound 53, Compound 54Compound 55, Compound 56 and Compound 57. General Procedures: Method F:

To a suspension ofDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (0.150 g, 0.44 mmol) in CH₂Cl₂ (4 mL) wasadded TEA (0.185 mL, 1.32 mmol) and the reaction mixture was stirred for10 min and cooled in an ice bath with continuous stirring. The acylchloride (0.484 mmol) was dissolved in CH₂Cl₂ (1 mL) and added dropwiseto the reaction mixture. The reaction mixture was allowed to reach roomtemperature, stirred for 16 h and partitioned with H₂O (3×4 mL), washedwith brine (3×4 mL), NaOH (0.5 M, 3×4 mL) and the organic layer wasevaporated, adsorbed on silica gel (0.3 g) with EtOAc. The desiredproduct was isolated by column chromatography using a gradient 0 to 8%[v/v] MeOH in EtOAc. The solid obtained was dissolved in ethanol (1 mL)and a solution of HCl (0.8 M, 1 mL) in EtOH was added. Evaporation ofthe volatiles led to the corresponding hydrochloride salt.

N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)hexanamidehydrochloride Compound 51

The compound was prepared according to method F with hexanoyl chloride(59 mg, 0.484 mmol) andDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22.N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)hexanamidehydrochloride Compound 51 was obtained as an off white solid. (56 mg,34% yield).

MW: 369.89; Yield: 34%; Off White Solid; Mp (° C.): 182.0.

¹H-NMR (CD₃OD, *): 0.84 (t, 3H, J=6.7, CH₃), 1.10-1.32 (m, 4H, CH₂),1.35-1.50 (m, 2H, CH₂), 1.80-2.00 (m, 4H, CH₂), 2.05-2.30 (m, 2H, CH₂),3.35-3.45 (m, 2H, CH₂), 3.50-3.65 (m, 2H, CH₂), 5.09 (d, 1H, J=3.7 Hz,N—CH), 5.38 (d, 1H, J=3.7 Hz, O—CH), 8.14 (d, 2H, J=6.3 Hz, ArH), 8.80(d, 2H, J=6.3 Hz, ArH).

MS-ESI m/z (% rel. int.): 334.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 214 nm, Compound 51 RT=3.90 min, peak area99.0%.

N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)heptanamidehydrochloride Compound 52

The compound was prepared according to method F with heptanoyl chloride(72 mg, 0.484 mmol) andDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22.N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)heptanamidehydrochloride Compound 52 was obtained as an off white solid. (192 mg,66% yield).

MW: 383.91; Yield: 66%; Off White Solid; Mp (° C.): 187.1.

¹H-NMR (CD₃OD, *): 0.88 (t, 3H, J=3.7 Hz, CH₃), 1.15-1.37 (m, 6H, CH₂),1.37 (m, 2H, CH₂), 1.85-2.02 (m, 4H, CH₂), 1.18-2.27 (m, 2H, CH₂),3.37-3.50 (m, 2H, N—CH₂), 3.55-3.70 (m, 2H, NCH₂), 5.14 (d, 1H, N—CH),5.42 (d, 1H, O—CH), 8.19 (d, 2H, J=6.3 Hz, ArH), 8.83 (d, 2H, J=6.3 Hz,ArH).

¹³C-NMR (CD₃OD, *): 14.4, 23.6, 25.0, 26.7, 27.0, 29.9, 32.6, 36.4,47.5, 56.7, 72.6, 126.6, 142.0, 164.5, 169.2, 175.9.

MS-ESI m/z (% rel. Int.): 348.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 254 nm, Compound 52 RT=4.10 min, peak area99.0%.

N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)octanamidehydrochloride Compound 53

The compound was prepared according to method F with octanoyl chloride(78 mg, 0.484 mmol) andDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22.N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)octanamidehydrochloride Compound 53 was obtained as an off white solid. (131 mg,75% yield).

MW: 397.94; Yield: 75%; Off White Solid; Mp (° C.): 185.9.

¹H-NMR (CD₃OD, *): 0.91 (t, 3H, J=6.4 Hz, CH₃), 1.12-1.37 (m, 8H, CH₂),1.40-1.52 (m, 2H, CH₂), 1.81 (m, 4H, CH₂), 2.12-2.25 (m, 2H, CH₂),3.40-3.52 (m, 2H, N—CH₂), 3.55-3.65 (m, 2H, N—CH₂), 5.14 (d, 1H, J=3.7Hz, N—CH), 5.43 (d, 1H, J=3.7 Hz, OCH), 8.19 (d, 2H, J=6.3 Hz, ArH),8.84 (d, 2H, J=6.3 Hz, ArH).

¹³C-NMR (CD₃OD, *): 14.4, 23.7, 24.9, 25.0, 25.5, 26.8, 27.0, 30.1,30.2, 32.8, 34.3, 36.5, 47.5, 56.8, 71.6, 72.6, 126.7, 126.9, 142.1,143.7, 164.5, 169.2, 176.0.

MS-ESI m/z (% rel. Int.): 362.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 254 nm, Compound 53 RT=4.37 min, peak area99.9%.

N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)palmitamidehydrochloride Compound 54

The compound was prepared according to method F with palmitoyl chloride(133 mg, 0.484 mmol) andDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22.N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)palmitamidehydrochloride Compound 54 was obtained as an off white solid. (105 mg,47% yield).

MW: 510.15; Yield: 47%; White Solid; Mp (° C.): 185.9.

¹H-NMR (CD₃OD, *): 0.92 (t, 3H, CH₃), 1.18-1.42 (m, 24H, CH₂), 1.42-1.58(m, 2H, CH₂), 1.85 (m, 4H, CH₂), 2.15 (m, 2H, CH₂), 3.41-3.50 (m, 2H,CH₂), 3.50-3.68 (m, 2H, CH₂), 5.14 (d, 1H, J=3.5 Hz, N—CH), 5.42 (d, 1H,J=3.5 Hz, O—CH), 8.18 (d, 2H, J=6.0 Hz, ArH), 8.82 (d, 2H, J=5.7 Hz,ArH).

¹³C-NMR (CD₃OD, *): 14.4, 23.7, 25.0, 26.8, 27.0, 30.3, 30.4, 30.5,30.6, 30.7, 30.8, 33.1, 36.4, 47.5, 56.8, 72.6, 126.6, 142.1, 164.5,169.2, 175.9.

MS-ESI m/z (% rel. Int.): 474.2 ([MH]⁺, 40).

HPLC: Method A, detection UV 254 nm, Compound 54 RT=6.36 min, peak area97.0%.

N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)benzamidehydrochloride Compound 55

The compound was prepared according to method F with benzoyl chloride(141 mg, 0.484 mmol) andDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22.N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)benzamidehydrochloride Compound 55 was obtained as an off white solid. (67 mg,34% yield).

MW: 375.85; Yield: 34%; Off White Solid; Mp (° C.): 212.

¹H-NMR (CD₃OD, *): 1.69-1.91 (m, 4H, CH₂), 3.25-3.40 (m, 2H, N—CH₂),3.40-3.58 (m, 2H, N—CH₂), 5.22 (d, 1H, J=3.7 Hz, N—CH), 5.43 (d, 1H,J=3.5 Hz, O—CH), 7.32 (t, 2H, J=7.8 Hz, ArH), 7.40 (t, 1H, J=6.9 Hz,ArH), 7.63 (d, 2H, J=7.1 Hz, ArH), 8.08 (d, 2H, J=6.6 Hz, ArH), 8.66 (d,2H, J=6.1 Hz, ArH).

¹³C-NMR (CD₃OD, *): 25.0, 27.1, 47.6, 57.6, 72.7, 126.6, 128.4, 129.7,133.3, 134.4, 142.1, 164.5, 169.0, 169.7.

MS-ESI m/z (% rel. Int.): 340.2 ([MH]⁺, 5).

HPLC: Method A, detection UV 254 nm, Compound 55 RT=3.66 min, peak area99.0%.

N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)-4-methoxy-benzamidehydrochloride Compound 56

The compound was prepared according to method F with 4-methoxybenzoylchloride (82 mg, 0.484 mmol) andDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22.N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)-4-methoxy-benzamidehydrochloride Compound 56 was obtained as an off white solid. (105 mg,58% yield).

MW: 405.87; Yield: 58%; Off White Solid; Mp(° C.): 205.3 (dec).

¹H-NMR (CD₃OD, *): 1.82-2.08 (m, 4H, CH₂), 3.45-3.55 (m, 2H, CH₂—N),3.60-3.70 (m, 2H, NCH₂), 3.86 (s, 3H, O—CH₃), 5.35 (d, 1H, J=3.7 Hz,N—CH), 5.56 (d, 1H, J=3.6 Hz, O—CH), 6.99 (dd, 2H, J=6.9 Hz, J=1.9 Hz),7.76 (dd, 2H, J=6.9 Hz, J=1.9 Hz, ArH), 8.21 (d, 2H, J=6.6 Hz, ArH),8.79 (d, 2H, J=6.6 Hz, ArH).

¹³C-NMR (CD₃OD, *): 25.0, 27.1, 47.6, 56.0, 57.5, 72.7, 114.9, 115.2,126.3, 126.6, 130.4, 133.7, 142.1, 164.4, 164.5, 169.1.

MS-ESI m/z (% rel. Int.): 370.2 ([MH]⁺, 10).

HPLC: Method A, detection UV 254 nm, Compound 56 RT=3.76 min, peak area99%.

3,4-Dichloro-N-(DL-threo-1-Hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)benzamideCompound 57

The compound was prepared according to method F with 3,4-dichlorobenzoylchloride (101 mg, 0.484 mmol) andDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydro chloride Compound 22.3,4-Dichloro-N-(DL-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)benzamideCompound 57 was obtained as an off white solid. (92 mg, 55% yield).

MW: 444.74; Yield: 55%; Off White Solid; Mp (° C.): 319.5 (dec).

¹H-NMR (CD₃OD, *): 1.82-2.05 (m, 4H, CH₂), 3.40-3.70 (m, 4H, N—CH₂),5.33 (d, 1H, J=3.9 Hz, N—CH), 5.55 (d, 1H, J=4.0 Hz, O—CH), 7.61-7.75(m, 2H, ArH), 7.96 (d, 1H, J=1.5 Hz, ArH), 8.22 (d, 2H, J=6.4 Hz, ArH),8.81 (d, 2H, J=6.0 Hz, ArH).

¹³C-NMR (CD₃OD, *): 25.0, 27.0, 57.8, 72.6, 126.6, 128.3, 130.7, 131.9,133.8, 134.7, 137.2, 142.2, 164.3, 167.2, 168.8.

MS-ESI m/z (% rel. Int.): 408.0, ([MH]⁺, 10)

HPLC: Method A, detection UV 254 nm, Compound 57 RT=4.28 min, peak area99.9%.

Preparation of Compound 58, Compound 59, Compound 60, Compound 61,Compound 62, Compound 63, Compound 64, Compound 65, Compound 66,Compound 67, Compound 68, Compound 69. General Procedures: Method G (inCH₂Cl₂):

To a stirred solution ofDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (0.15 g, 0.49 mmol) in 10 mL of CH₂Cl₂ at+4° C. were added triethylamine (200 μl, 1.45 mmol) and very slowly acidchloride in 3 mL of CH₂Cl₂. The mixture was stirred overnight at RTunder nitrogen and then partitioned between CH₂Cl₂ and 1 N aqueoussodium carbonate. The organic layer was evaporated and the obtainedresidue purified by column chromatography on silica (EtOAc:MeOH=95:5).The hydrochloride salt was obtained in MeOH at 0° C. with 0.3 M HCl indiethylether to give after evaporation of solvents and drying theacylated compound.

Method H (in MeOH):

To a stirred solution ofDL-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (0.20 g, 0.65 mmol) in 3 mL of MeOH wereadded triethylamine (180 μl, 1.30 mmol) and aldehyde (or ketone). Themixture was stirred overnight at RT under nitrogen and then was addedAcOH (200 μL, 3.2 mmol) and NaBH₃CN. After 5 h at 20° C., MeOH wasevaporated and the residue was partitioned between CH₂Cl₂ and 1 Naqueous sodium carbonate. The organic layer was evaporated and theobtained residue was purified by column chromatography on silica(EtOAc:MeOH or CH₂Cl₂:MeOH). The hydrochloride salt was obtained in MeOHat 0° C. with 0.3 M HCl in diethylether to give after evaporation ofsolvents and drying the alkylated compound.

BenzylDL-threo-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamatehydrochloride Compound 58

The compound was prepared according to method G with benzylchloroformate (91 mg, 0.53 mmol). After work-up benzylDL-threo-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamatehydrochloride Compound 58 was obtained as a white solid (90 mg, 46%yield).

MW: 405.9; Yield: 46.0%; White Solid; Mp (° C.): 185.3.

R_(f): 0.38 (MeOH:EtOAc=10:90) free base.

¹H-NMR (CD₃OD, *): 1.87-2.03 (m, 4H, 2×CH₂), 3.40-3.48 (m, 2H, CH₂N),3.56-3.62 (m, 2H, CH₂N), 4.85-5.04 (m, 3H, CH₂O, CHO), 5.39 (d, 1H,J=2.8 Hz, NH), 7.26-7.36 (m, 5H, ArH), 8.12 (d, 2H, J=6.0 Hz, ArH), 8.69(d, 2H, J=6.0 Hz, ArH).

¹³C-NMR (CD₃OD, *): 25.0, 27.0, 47.5, 48.0, 58.8, 67.9, 72.7, 126.6(2×C), 129.1, 129.2, 129.5, 138.1, 141.9 (2×C), 158.1, 164.4, 169.2.

MS-ESI m/z (% rel. Int.): 370.1 ([MH]⁺, 15), 219.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 58 RT=4.10 min, peak area99.8%.

N-(DL-threo-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-yl)decanamidehydrochloride Compound 59

The compound was prepared according to method G with decanoyl chloride(111 μL, 0.53 mmol). After work-upN-(DL-threo-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-yl)decanamidehydrochloride Compound 59 was obtained as a white solid (115 mg, 55%yield).

MW: 425.99; Yield: 55%; White Solid; Mp (° C.): 184.8.

R_(f): 0.22 (MeOH:EtOAc=5:95) free base.

¹H-NMR (CD₃OD, *): 0.90 (t, 3H, J=7.0 Hz, CH₃), 1.26-1.34 (m, 12H,6×CH₂), 1.42-1.50 (m, 2H, CH₂), 1.86-1.98 (m, 4H, 2×CH₂), 2.13-2.20 (m,2H, CH₂CO), 3.41-3.46 (m, 2H, CH₂N), 3.52-3.61 (m, 2H, CH₂N), 5.12 (d,1H, J=3.8 Hz, CH), 5.40 (d, 1H, J=3.7 Hz, CH), 8.16 (d, 2H, J=6.5 Hz,ArH), 8.97 (d, 2H, J=6.7 Hz, ArH).

¹³C-NMR (CD₃OD, *): 14.4, 23.7, 25.0, 26.8, 27.0, 30.3, 30.4, 30.6,33.0, 36.5, 47.5, 56.8, 72.6, 126.6 (2×C), 142.1 (2×C), 164.4, 169.2,175.9.

MS-ESI m/z (% rel. Int.): 390.1 ([MH]⁺, 20), 219.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 59 RT=4.9 min, peak area99.5%.

DL-threo-2-(Benzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 60

The compound was prepared according to method H with benzaldehyde (78mg, 0.72 mmol). After column chromatography (EtOAc:MeOH=95:5) and HCltreatmentDL-threo-2-(benzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 60 was obtained as a white solid (114 mg, 46%yield).

MW: 398.33; Yield: 46%; White Solid; Mp (° C.): 131.5.

R_(f): 0.60 (MeOH:EtOAc=10:90) free base.

¹H-NMR (CD₃OD, *): 1.35-1.72 (m, 4H, 2×CH₂), 2.10-2.18 (m, 1H, CH₂N),2.78-2.86 (m, 1H, CH₂N), 3.18-3.24 (m, 2H, CH₂N), 4.22 (d, 1H, J=8.5 Hz,CH), 4.26-4.36 (m, 2H, CH₂N), 5.18 (d, 1H, J=8.5 Hz, CH), 7.43-7.51 (m,5H, BzH), 7.86 (d, 2H, J=6.6 Hz, ArH), 8.69 (d, 2H, J=6.6 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.7, 26.3, 47.3, 47.9, 51.4, 63.9, 72.4, 126.2,130.3, 131.0, 131.4, 131.5, 148.9, 156.1, 163.9.

MS-ESI m/z (% rel. Int.): 326.1 ([MH]⁺, 100), 227.0 (80).

HPLC: Method A, detection UV 254 nm, Compound 60 RT=4.30 min, peak area98.2%.

DL-threo-3-Hydroxy-2-(methylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 61

The compound was prepared according to method H with paraformaldehyde(21 mg, 0.65 mmol). After column chromatography (EtOAc:MeOH=7:3) and HCltreatmentDL-threo-3-hydroxy-2-(methylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 61 was obtained as a pale yellow solid (28 mg,13% yield).

MW: 322.23; Yield: 13%; Pale Yellow Solid.

R_(f): 0.20 (MeOH:EtOAc=30:70) free base.

¹H-NMR (CD₃OD, *): 1.60-1.80 (m, 4H, 2×CH₂), 2.61 (s, 1H, CH₃),2.68-2.76 (m, 1H, CH₂N), 3.24-3.57 (m, 3H, CH₂N), 4.53 (d, 1H, J=6.8 Hz,CH), 5.26 (d, 1H, J=7.0 Hz, CH), 8.11 (d, 2H, J=5.8 Hz, ArH), 8.85 (d,2H, J=5.6 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.8, 26.7, 32.8, 47.7, 48.3, 65.4, 71.7, 126.7(2×C), 143.4 (2×C), 161.1, 163.9.

MS-ESI m/z (% rel. Int.): 251.1 ([MH]⁺, 10), 151.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 61 RT=0.70 min, peak area97.5%.

DL-threo-3-Hydroxy-2-(pentylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 62

The compound was prepared according to method H with valeraldehyde (60mg, 0.68 mmol). After column chromatography (EtOAc:MeOH=95:5) and HCltreatmentDL-threo-3-hydroxy-2-(pentylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 62 was obtained as a white solid (107 mg, 44%yield).

MW: 378.34; Yield: 44%; White Solid; Mp (° C.): 86.6.

R_(f): 0.30 (MeOH:EtOAc=20:80) free base.

¹H-NMR (CD₃OD, *): 0.95 (t, 3H, J=6.4 Hz, CH₃), 1.32-1.40 (m, 6H,3×CH₂), 1.70-1.87 (m, 4H, 2×CH₂), 2.70-2.75 (m, 1H, CH₂N), 2.90-3.00 (m,1H, CH₂N), 3.10-3.39 (m, 3H, CH₂N), 3.46-3.60 (m, 1H, CH₂N), 4.61 (d,1H, J=7.5 Hz, CH), 5.39 (d, 1H, J=7.5 Hz, CH), 8.22 (d, 2H, J=6.2 Hz,ArH), 8.97 (d, 2H, J=6.2 Hz, ArH).

¹³C-NMR (CD₃OD, *): 9.3, 14.1, 23.2, 24.8, 26.7, 26.8, 29.7, 47.6, 47.9,64.5, 72.1, 126.8 (2×C), 143.3 (2×C), 161.1, 164.1.

MS-ESI m/z (% rel. Int.): 306.3 ([MH]⁺, 15), 207.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 62 RT=3.60 min, peak area98.5%.

DL-threo-3-Hydroxy-2-(hexylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 63

The compound was prepared according to method H with hexanal (71 mg,0.68 mmol). After column chromatography (EtOAc:MeOH=95:5) and HCltreatmentDL-threo-3-hydroxy-2-(hexylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 63 was obtained as a beige solid (112 mg, 45%yield).

MW: 392.36; Yield: 45%; Beige Solid; Mp (° C.): 108.2.

R_(f): 0.35 (MeOH:EtOAc=20:80) free base.

¹H-NMR (CD₃OD, *): 0.94 (t, 3H, J=7.2 Hz, CH₃), 1.30-1.42 (m, 6H,3×CH₂), 1.66-1.90 (m, 6H, 3×CH₂), 2.68-2.74 (m, 1H, CH₂N), 2.90-2.99 (m,1H, CH₂N), 3.09-3.16 (m, 1H, CH₂N), 3.32-3.39 (m, 1H, CH₂N), 3.47-3.60(m, 2H, CH₂N), 4.60 (d, 1H, J=7.7 Hz, CH), 5.38 (d, 1H, J=7.7 Hz, CH),8.24 (s, 2H, ArH), 8.97 (s, 2H, ArH).

¹³C-NMR (CD₃OD, *): 14.3, 23.4, 24.8, 26.7, 27.1, 27.2, 32.4, 47.6,64.4, 72.1, 126.9 (2×C), 143.2 (2×C), 161.3, 164.1.

MS-ESI m/z (% rel. Int.): 320.1 ([MH]⁺, 30), 221.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 63 RT=3.80 min, peak area97.8%.

DL-threo-3-Hydroxy-2-(heptylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 64

The compound was prepared according to method H with heptaldehyde (82mg, 0.68 mmol). After column chromatography with EtOAc:MeOH=95:5 and HCltreatmentDL-threo-3-hydroxy-2-(heptylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 64 was obtained as a white solid (121 mg, 47%yield).

MW: 406.39; Yield: 47%; White Solid; Mp (° C.): 242.4.

R_(f): 0.40 (MeOH:EtOAc=20:80) free base.

¹H-NMR (CD₃OD, *): 0.92 (t, 3H, J=7.0 Hz, CH₃), 1.32-1.40 (m, 8H,4×CH₂), 1.65-1.90 (m, 6H, 3×CH₂), 2.71-2.76 (m, 1H, CH₂N), 2.90-2.99 (m,1H, CH₂N), 3.09-3.38 (m, 1H, CH₂N), 3.47-3.62 (m, 2H, CH₂N), 4.61 (d,1H, J=7.5 Hz, CH), 5.39 (d, 1H, J=7.5 Hz, CH), 8.24 (d, 2H, J=6.0 Hz,ArH), 8.97 (d, 2H, J=5.9 Hz, ArH).

¹³C-NMR (CD₃OD, *): 14.4, 23.6, 24.8, 26.7, 27.1, 27.5, 29.9, 32.7,47.7, 64.4, 72.1, 126.9 (2×C), 143.2 (2×C), 161.4, 164.1.

MS-ESI m/z (% rel. Int.): 334.1 ([MH]⁺, 45), 235.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 64 RT=4.00 min, peak area97.5%.

DL-threo-2-(4-Methylbenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 65

The compound was prepared according to method H with4-methylbenzaldehyde (86 mg, 0.70 mmol). After column chromatography(EtOAc:MeOH=95:5) and HCl treatmentDL-threo-2-(4-methylbenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 65 was obtained as a white solid (137 mg, 51% yield).

MW: 412.35; Yield: 51%; White Solid; Mp (° C.): 87.5.

R_(f): 0.20 (MeOH:EtOAc=5:95) free base.

¹H-NMR (CD₃OD, *): 1.49-2.12 (m, 4H, 2×CH₂), 2.30-2.40 (m, 1H, CH₂N),2.35. (s, 3H, CH₃), 2.75-2.95 (m, 1H, CH₂N), 3.18-3.25 (m, 2H, CH₂N),4.12-4.32 (m, 3H, CH₂N, CH), 5.30 (d, 1H, J=7.9 Hz, CH), 7.24-7.39 (m,4H, BzH), 8.11 (d, 2H, J=6.7 Hz, ArH), 8.89 (d, 2H, J=6.6 Hz, ArH).

¹³C-NMR (CD₃OD, *): 21.2, 24.7, 26.3, 26.4, 47.4, 47.9, 51.2, 63.4,72.4, 126.1, 126.6 (2×C), 128.2, 130.9, 131.4, 131.5, 141.4, 143.5(2×C), 148.9, 156.1; 160.5, 163.8.

MS-ESI m/z (% rel. Int.): 340.1 ([MH]⁺, 10), 104.9 (100).

HPLC: Method A, detection UV 254 nm, Compound 65 RT=3.70 min, peak area97.3%.

DL-threo-2-(4-Chlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 66

The compound was prepared according to method H with4-chlorobenzaldehyde (98 mg, 0.70 mmol). After column chromatography(EtOAc:MeOH=95:5) and HCl treatmentDL-threo-2-(4-chlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 66 was obtained as a white solid (126 mg, 45% yield).

MW: 432.77; Yield: 45%; White Solid; Mp (° C.): 122.7.

R_(f): 0.20 (MeOH:EtOAc=5:95) free base.

¹H-NMR (CD₃OD, *): 1.54-1.75 (m, 4H, 2×CH₂), 2.41-2.49 (m, 1H, CH₂N),3.08-3.15 (m, 1H, CH₂N), 3.19-3.27 (m, 2H, CH₂N), 4.25-4.47 (m, 3H,CH₂N, CH), 5.34 (d, 1H, J=7.9 Hz, CH), 7.46-7.55 (m, 4H, BzH), 8.15 (d,2H, J=6.0 Hz, ArH), 8.92 (d, 2H, J=5.7 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.7, 26.5, 47.5, 47.8, 50.8, 63.8, 72.3, 126.7(2×C), 130.3, 133.4, 137.1, 143.5 (2×C), 149.0, 160.6, 163.8.

MS-ESI m/z (% rel. Int.): 360.1/362.1 ([MH]⁺, 20), 124.9 (100).

HPLC: Method A, detection UV 254 nm, Compound 66 RT=3.70 min, peak area97.0%.

DL-threo-2-(4-Methoxybenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneone Compound 67

The compound was prepared according to method H with4-methoxybenzaldehyde (95 mg, 0.70 mmol). After column chromatography(EtOAc:MeOH=95:5) and HC1 treatmentDL-threo-2-(4-methoxybenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneone Compound 67 was obtained as a white solid (123 mg, 44% yield).

MW: 428.35; Yield: 44%; White Solid; Mp (° C.): 193.2.

R_(f): 0.20 (MeOH:EtOAc=5:95) free base.

¹H-NMR (CD₃OD, *): 1.51-1.72 (m, 4H, 2×CH₂), 2.38-2.41 (m, 1H, CH₂N),2.94-3.01 (m, 1H, CH₂N), 3.18-3.28 (m, 2H, CH₂N), 3.81 (s, 3H, CH₃O),4.18-4.34 (m, 3H, CH₂N, CH), 5.31 (d, 1H, J=7.3 Hz, CH), 6.97 (d, 2H,J=8.5 Hz, ArH), 7.42 (d, 2H, J=8.5 Hz, ArH), 8.13 (d, 2H, J=6.5 Hz,ArH), 8.91 (d, 2H, J=6.3 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.7, 26.5, 47.5, 47.9, 51.1, 55.9, 63.2, 72.3,115.5, 122.9, 126.8 (2×C), 133.1, 143.3 (2×C), 160.9, 162.4, 163.8.

MS-ESI m/z (% rel. Int.): 356.1 ([MH]⁺, 10), 120.9 (100).

HPLC: Method A, detection UV 254 nm, Compound 67 RT=3.50 min, peak area98.6%.

DL-threo-2-(3,4-Dichlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 68

The compound was prepared according to method H with3,4-dichlorobenzaldehyde (122 mg, 0.70 mmol). After columnchromatography (EtOAc:MeOH=95:5) and HCl treatmentDL-threo-2-(3,4-dichlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 68 was obtained as a white solid (153 mg, 50%yield).

MW: 467.22; Yield: 50%; White Solid; Mp (° C.): 190.3.

R_(f): 0.20 (MeOH:EtOAc=5:95) free base.

¹H-NMR (CD₃OD, *): 1.58-1.76 (m, 4H, 2×CH₂), 2.48-2.55 (m, 1H, CH₂N),2.85-3.00 (m, 1H, CH₂N), 3.18-3.26 (m, 2H, CH₂N), 4.23-4.41 (m, 2H,CH₂N), 4.54 (d, 1H, J=7.7 Hz, CH), 5.34 (d, 1H, J=7.2 Hz, CH), 7.48 (d,1H, J=8.3 Hz, ArH), 7.63 (dd, 1H, J=8.2 Hz, J=1.4 Hz, ArH), 7.75 (s, 1H,ArH), 8.16 (d, 2H, J=5.4 Hz, ArH), 8.92 (d, 2H, J=5.3 Hz, ArH).

¹³C-NMR (CD₃OD, *): 23.2, 25.0, 46.0, 46.4, 48.9, 62.5, 70.7, 125.3,130.2, 130.6, 130.7, 132.3, 133.6, 141.9, 159.2, 162.3.

MS-ESI m/z (% rel. Int.): 394.1/396.1 ([MH]⁺, 40), 110.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 68 RT=3.90 min, peak area99.0%.

DL-threo-2-(4-Methoxybenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 69

The compound was prepared according to method H with cyclohexanone (75μL, 0.70 mmol). After column chromatography (EtOAc:MeOH=95:5) and HCltreatmentDL-threo-2-(4-methoxybenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 69 as a white solid (154 mg, 61% yield).

MW: 390.35; Yield: 61%; White Solid; Mp (° C.): 144.1.

R_(f): 0.25 (MeOH:EtOAc=5:95) free base.

¹H-NMR (CD₃OD, *): 1.16-2.15 (m, 15H, 7×CH₂, CH), 2.65-2.72 (m, 1H,CH₂N), 3.07-3.15 (m, 1H, CH₂N), 3.43-3.65 (m, 2H, CH₂N), 4.61 (d, 1H,J=7.8 Hz, CH—N), 5.35 (d, 1H, J=7.8 Hz, CH—O), 8.21 (d, 2H, J=6.3 Hz,ArH), 8.95 (d, 2H, J=6.1 Hz, ArH).

¹³C-NMR (CD₃OD, *): 23.2, 24.0, 24.2, 24.4, 25.2, 28.2, 29.3, 46.1,57.4, 60.5, 71.0, 125.4 (2×C), 141.8 (2×C), 159.5, 162.6.

MS-ESI m/z (% rel. Int.): 318.2 ([MH]⁺, 40; 219.1, 100).

HPLC: Method A, detection UV 254 nm, Compound 69 RT=3.40 min, peak area99.7%.

Preparation of(±)-threo-2-amino-3-(furan-2-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 201trans-5-(Furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136D

BLE 04136D was prepared in accordance with method D using 2-furaldehyde(0.449 mL, 5.42 mmol), KOH (0.276 mg, 4.92 mmol) in methanol (5 mL) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.75 g, 5.42 mmol).After work-up the residue was purified by column chromatography (SiO₂,cyclohexane:EtOAc=100:0 to 0:100) to led, after evaporation, totrans-5-(furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04138D (0.742 g, 58.5% yield) as a pale yellow oil.

MW: 234.25; Yield: 58.5%; Pale Yellow Oil.

¹H-NMR (CDCl₃,*): 1.80-2.10 (m, 4H, CH₂), 3.47-3.60 (m, 3H, CH₂N),3.93-4.03 (m, 1H, CH₂N), 4.94 (dd, 1H, J=7.4 Hz, J=2.2 Hz, CH—N), 6.14(d, 1H, J=7.4 Hz, CH—O), 6.37 (dd, 1H, J=3.3 Hz, J=1.8 Hz, CH═C), 6.47(d, 1H, J=3.3 Hz, CH═C), 6.92 (d, 1H, J=2.2 Hz, O—CH═N), 7.44 (t, 1H,J=1.6 Hz, OCH═C).

(±)-threo-2-Amino-3-(furan-2-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 201

Compound 201 was prepared following method E withtrans-(5-(furan-2-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneBLE 04136D (0.30 g, 1.28 mmol), hydrochloric acid 37% (0.3 mL) andmethanol (10 mL). After overnight at RT and work-up(±)-threo-2-amino-3-(furan-2-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onehydrochloride (0.22 g, 66% yield) was obtained as a pale brown solid.

MW: 260.72; Yield: 66%; Pale Brown Solid; Mp (° C.): 159.8

¹H-NMR (CD₃OD,*): 1.62-1.95 (m, 4H, 2×CH₂), 2.72-2.85 (m, 1H, CH₂N),3.22-3.35 (m, 1H, CH₂N), 3.38-3.55 (m, 2H, CH₂N), 4.35 (d, 1H, J=8.5 Hz,CH—N), 4.91 (d, 1H, J=8.5 Hz, CH—O), 6.45 (m, 2H, ArH), 7.52-7.57 (m,1H, ArH).

¹³C-NMR (CD₃OD,*): 24.9, 26.9, 47.4, 47.6, 57.4, 67.8, 109.9, 111.9,144.4, 153.0, 166.0.

MS-ESI m/z (% rel. Int.): 225.1 ([MH]⁺, 18), 207.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 201 RT=2.87 min, peak area92.0%.

Preparation of(−)-(2R,3S)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 203 and(+)-(2S,3R)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 204 Extraction of the Free Base of Compound 22:

(±)-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (350 mg, 1.14 mmol) was dissolved in 20 mLof a K₂CO₃ (10%) solution and the aqueous mixture was then saturatedwith NaCl. The aqueous phase was extracted by a mixtureCH₂Cl₂:2-PrOH=9:1 (6×15 mL). The organic phase was dried over MgSO₄ andevaporated to afford 226 mg (85% yield) of the free base of Compound 22.

Analytical Chiral Separation:

20 μL of a 1 mg/mL solution of Compound 22 were injected on ChiralpakAD: flow-rate=1 mL/min, temperature=25° C., mobile phase:hexane:ethanol=1:1, detection by UV at 220 nm and by polarimeter, Rt(−)=8.20 min, Rt (+)=10.61 min, k (−)=1.72, k (+)=2.51, α=1.47 andresolution Rs=3.08.

Semi-Preparative Chiral Separation:

A solution of 100 mg/mL was prepared and 10 μL of this solution wereinjected every 4.5 min on Chiralpak AD, flow-rate=1 mL/min, mobile phasehexane:ethanol=4:6, detection by UW at 254 nm. 135 successive injectionswere done. The two main fractions were identified by UW and collected intwo different flasks. The solvent was removed in vacuo at 30° C. Theresulting solid was dissolved in 50 mL of CH₂Cl₂ and then filtered on a0.45 μm millipore membrane. After evaporation of CH₂Cl₂, the solid wasdissolved in 50 mL of methanol and then filtered. The salts wereregenerated according to the procedure reported above.

Regeneration of the Salt:

After the chiral separation, about 63 mg of each enantiomer of the freebase were dissolved in 100 mL of ethanol and 6.7 mL of HCl (0.2 N, 5 eq)were added. The solvent was evaporated, then 50 mL of ethanol were addedand then removed in vacuo and the products were dried over P₂O₅ undervacuum overnight. The enantiomeric purity of the products was checked byanalytical injection of the regenerated salts:

(−)-(2R,3S)-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 203

MW: 308.20; 83 mg obtained; Yield: 23.5%; White Solid; Mp (° C.): 183.5

Enantiomeric excess=99.3%

α²⁵ _(D)=−22.7 (MeOH, c=0.51).

(+)-(2S,3R)-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 204

MW: 308.20; 82 mg obtained; Yield: 23.5%; White Solid; Mp (° C.): 176.9

Enantiomeric excess=98.5%

α²⁵ _(D)=+23.1 (MeOH, c=1).

Preparation of(+)-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 205 and(−)-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 206 Extraction of the Free Base:

(±)-threo-2-amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 23 (243 mg, 0.88 mmol) was dissolved in 10 mL ofa Na₂CO₃ (10%) solution and the aqueous mixture was then saturated withNaCl. The aqueous phase was extracted by 5×15 mL of a mixtureCH₂Cl₂:2-PrOH=9:1. The organic phase was dried over MgSO₄ and evaporatedto afford 190 mg (90%) of the free base of Compound 23.

Analytical Chiral Separation:

20 μL of a 1 mg/mL solution of Compound 23 were injected on ananalytical Chiralpak AD: flow-rate=1 mL/min, temperature=25° C., mobilephase: ethanol, detection by UW at 220 nm and by polarimeter, Rt(+)=4.98 min, Rt (−)=6.23 min, k (+)=0.55, k (−)=0.93, α=1.17 andresolution Rs=3.34.

Regeneration of the Salt:

After the chiral separation, about 70 mg of each enantiomer of the freebase were dissolved in 100 mL of ethanol and 3.6 mL of HCl (0.2 N, 2.5eq) were added. The solvent was evaporated then 50 mL of ethanol wereadded and then removed in vacuo. The product was dissolved in 2 mL ofmethanol and 3 mL of ethyl acetate were added. The solvents were removedto give a white solid and then, the solids were dried over P₂O₅ undervacuum overnight.

Semi-Preparative Chiral Separation:

A 175 mg/mL solution of the free base was prepared and 6 μL of thissolution were injected every 3 min on an analytical Chiralpak AD,flow-rate=1 mL/min, mobile phase ethanol, detection by UV at 254 nm. 150successive injections were done. The two main fractions were identifiedby UV and collected in two different flasks. The solvent was removed invacuo at 30° C. The resulting solid was dissolved in 50 mL of CH₂Cl₂ andthen filtered on a 0.45 μm millipore membrane. After evaporation ofCH₂Cl₂, the solid was dissolved in 50 mL of methanol and then filtered.The salt was regenerated according to the procedure reported above.

The enantiomeric purity of the products was checked by analyticalinjection of the regenerated salts:

(+)-threo-2-Amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 205

MW: 276.78; 83 mg obtained; Yield: 34%; White Solid; Mp (° C.): toohygroscopic.

Enantiomeric excess=99.5%

α²⁵ _(D)=+20.4 (MeOH, c=0.5).

(−)-threo-2-Amino-3-hydroxy-1-(pyrrolidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 206

MW: 276.78; 77 mg obtained; Yield: 32%; White Solid; Mp (° C.): toohygroscopic.

Enantiomeric excess=99.0%

α²⁵ _(D)=−20.0 (MeOH, c=0.52).

Preparation of(−)-threo-2-amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 207 and(+)-threo-2-amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 208 Semi-Preparative Separation was Performedon Chiralpak AD (250×10 mm): The Semi-Preparative Chiral SeparationNeeded Three Steps:

First step A: A 80 mg/mL solution of(±)-threo-2-amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 20 free base (220 mg) racemate was prepared and 200 μL of thissolution were injected every 8 min on Chiralpak AD, flow-rate=4 mL/min,mobile phase: ethanol, detection by UV at 290 nm. Two main fractionswere collected after 13 successive injections:

-   -   1A containing about 61 mg of (−) enantiomer Compound 207 free        base with ee>97%.    -   2A containing about 135 mg of mixture+/−enantiomers in a 74/26        ratio.

Second step B: A 30 mg/mL solution of fraction 2A was prepared and 100μL of this solution were injected every 6 min on Chiralpak AD,flow-rate=4 mL/min, mobile phase: ethanol, detection by UV at 290 nm.Two main fractions were collected after 45 successive injections:

-   -   1B containing about 27 mg of (−) enantiomer Compound 207 free        base withee>97%.    -   2B containing about 105 mg of mixture (+)/(−) in a 93/7 ratio.

Third step C: A 15 mg/mL solution of fraction 2B was prepared and 250 μLof this solution were injected every 6 min on Chiralpak AD, flow-rate=4mL/min, mobile phase: ethanol, detection by UV at 254 nm. Two mainfractions were collected after 28 successive injections:

-   -   1C containing about 7 mg of (−) enantiomer Compound 207 free        base with ee>97%.    -   2C containing about 89 mg of (+) enantiomer with ee>97%.

Fractions 1A, 1B and 1C of (−) enantiomer Compound 207 free base weremixed together. Fraction 2C of (+) enantiomer Compound 208 free base wastaken alone. For the both enantiomers, the solvent was removed in vacuoat 30° C. The resulting solid was dissolved in 50 mL of CH₂Cl₂ and thenfiltered on a 0.45 μm millipore membrane. After evaporation of CH₂Cl₂,the solid was dissolved in 50 mL of methanol and then filtered. The saltwas regenerated according to the procedure reported below. Theintermediate fraction collected contains 25 mg of a mixture of the bothenantiomers in 50/50 (+)/(−) ratio and some impurities.

Regeneration of the Salt (Dihydrochloride):

After the chiral separation, about 90-95 mg of each enantiomers of thefree base were dissolved in 100 mL of ethanol and 10 mL of HCl (0.2 N, 5eq) were added. The solvent was evaporated and 50 mL of ethanol wereadded and then removed in vacuo. The product was dissolved in 1 mL ofmethanol and 5 mL of ethyl acetate were added to precipitate the salt.The solvents were removed to give a white solid and then, the solidswere dried over P₂O₅ under vacuum overnight.

The enantiomeric purity of the products was checked by analytical HPLCinjection of the regenerated dihydrochloride salts:

(−)-threo-2-Amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 207

MW: 308.20; 124 mg obtained; Yield: 43%; White Solid; Mp (° C.): 120.4

Enantiomeric excess=97.8% measured by HPLC at 220 nm (Chiralpak AD)

RT=6.24 min, eluent ethanol, flow 1 mL/min.

α²⁵ _(D)=−15.9 (MeOH, c=1).

(+)-threo-2-Amino-3-hydroxy-3-(pyridin-3-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 208

MW: 308.20; 117 mg obtained; Yield: 40.5%; White Solid; Mp (° C.): 120.1

Enantiomeric excess=98.0% measured by HPLC at 220 nm (Chiralpak AD)

RT=7.39 min, eluent ethanol, flow 1 mL/min.

α²⁵ _(D)=+15.8 (MeOH, c=1).

Preparation of(±)-threo-2-amino-3-hydroxy-3-(2-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 209trans-(4,5-Dihydro-5-(2-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneVIB 01090A

VIB 01090A was prepared in accordance with method D using2-isocyano-1-(pyrrolidin-1-yl)ethanone SLA 09100 (327.9 mg, 2.155 mmol),potassium hydroxide (121 mg, 2.155 mmol) in methanol (2.2 mL) and2-iodo-benzaldehyde (500 mg, 2.155 mmol). The solution was stirred for 3h at 0° C. After work-up the crude product was purified by columnchromatography (florisil, EtOAc:MeOH=9:1) to obtain after evaporationtrans-(4,5-dihydro-5-(2-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneVIB 01090A as a yellow oil (364 mg, 44% yield).

MW: 384.22; Yield: 44%; Yellow Oil.

R_(f): 0.51 (EtOAc:MeOH=9:1).

¹H-NMR (CDCl₃, δ): 1.85-2.07 (m, 4H, 2×CH₂), 3.50-3.62 (m, 3H, CH₂),3.78-3.90 (m, 1H, CH₂), 4.57 (dd, 1H, J=5.6 Hz, J=1.9 Hz, CH—N), 6.19(d, 1H, J=5.6 Hz, CH—O), 7.05 (dt, 1H, J=7.7 Hz, J=1.6 Hz, ArH), 7.15(d, 1H, J=1.9 Hz, HC═N), 7.27 (dd, 1H, J=7.9 Hz, J=1.6 Hz, ArH), 7.39(t, 1H, J=7.3 Hz, ArH), 7.87 (d, 1H, J=7.8 Hz, ArH).

¹³C-NMR (CDCl₃, δ): 24.3, 26.0, 46.3, 46.6, 74.8, 84.4, 95.0, 126.5,128.4, 129.9, 139.8, 142.2, 155.7, 167.1.

(±)-threo-2-Amino-3-hydroxy-3-(2-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 209

Compound 209 was prepared following method E withtrans-(4,5-dihydro-5-(2-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneVIB 01090A (0.345 g, 0.89 mmol), HCl 37% (0.22 mL) and methanol (4 mL).After heating at 50° C. for 3 h and work-up, a trituration with EtOAcfollowed by filtration and drying afforded to(±)-threo-2-amino-3-hydroxy-3-(2-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 209 as a white solid (287 mg, 74% yield).

MW: 396.65; Yield: 74%; White Solid; Mp (° C.): 164.0

¹H-NMR (CD₃OD, δ): 1.47-1.90 (m, 4H, 2×CH₂), 1.95-2.10 (m, 1H, CH₂),3.25-3.55 (m, 3H, CH₂), 4.23 (d, 1H, J=9.0 Hz, CH—N), 5.20 (d, 1H, J=9.0Hz, CH—O), 7.11 (t, 1H, J=7.4 Hz, ArH), 7.50 (t, 1H, J=7.5 Hz, ArH),7.78 (d, 2H, J=7.9 Hz, ArH), 7.88 (d, 2H, J=7.7 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.8, 26.8, 47.8, 48.1, 58.6, 75.8, 98.3, 130.1,130.8, 131.8, 141.1, 143.8, 165.6.

MS-ESI m/z (% rel. Int.): 360.9 ([MH]⁺, 100), 342.9 (40).

HPLC: Method A, detection UV 254 nm, Compound 209 RT=3.88 min, peak area97.8%.

Preparation of(±)-threo-2-amino-3-hydroxy-3-hydroxy-3-(4-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 210trans-(4,5-Dihydro-5-(4-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneVIB 01090B

VIB 01090B was prepared in accordance with method D using2-isocyano-1-(pyrrolidin-1-yl)ethanone SLA 09100 (327.9 mg, 2.155 mmol),potassium hydroxide (121 mg, 2.155 mmol) in methanol (2.2 mL) and4-iodobenzaldehyde (500 mg, 2.155 mmol). The solution was stirred for 3h at 0° C. After work-up, the crude product was washed in a minimumamount of MeOH and filtered to obtain after dryingtrans-(4,5-dihydro-5-(4-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneVIB 01090B as a white solid (0.377 g, 52% yield).

MW: 384.22; Yield: 52%; White Solid; Mp (° C.): 115.1

¹H-NMR (CDCl₃, δ): 1.75-2.10 (m, 4H, 2×CH₂), 3.35-3.60 (m, 3H, CH₂),3.88-4.02 (m, 1H, CH₂), 4.53 (dd, 1H, J=7.7 Hz, J=1.9 Hz, CH—N), 6.09(d, 1H, J=7.7 Hz, CH—O), 6.92-7.11 (m, 3H, 2×ArH & CH═N), 7.69-7.70 (dd,2H, J=8.4 Hz, J=1.7 Hz, ArH).

¹³C-NMR (CDCl₃, δ): 24.1, 26.0, 46.2, 46.4, 75.7, 80.8, 94.0, 127.6(2×C), 137.9 (2×C), 139.4, 155.2, 166.4.

(±)-threo-2-Amino-3-hydroxy-3-hydroxy-3-(4-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 210

Compound 210 was prepared following method E withtrans-(4,5-dihydro-5-(4-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneVIB 01090B (0.345 g, 0.89 mmol), hydrochloric acid 37% (0.24 mL) andmethanol (4.4 mL). After heating at 50° C. for 3 h and work-up, atrituration with EtOAc followed by filtration and drying afforded to(±)-threo-2-amino-3-hydroxy-3-(4-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 210 as a white solid (247.5 mg, 64% yield).

MW: 396.65; Yield: 64%; White Solid; Mp (° C.): 184.4

¹H-NMR (CD₃OD, δ): 1.35-1.9 (m, 4H, 2×CH₂), 2.20-2.33 (m, 1H, CH₂),3.18-3.40 (m, 3H, CH₂), 4.11 (d, 1H, J=8.9 Hz, CH—N), 4.82 (d, 1H, J=8.9Hz, CH—O), 7.22 (d, 2H, J=8.2 Hz, ArH), 7.76 (d, 2H, J=8.2 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.8, 26.6, 47.3, 47.7, 59.2, 73.6, 95.1, 129.8(2×C), 138.9 (2×C), 140.6, 166.1.

MS-ESI m/z (% rel. Int.): 360.9 ([MH]⁺, 100), 342.9 (85).

HPLC: Method A, detection UV 254 nm, Compound 210 RT=4.08 min, peak area96.8%.

Preparation of(±)-threo-2-amino-3-hydroxy-3-(3-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 211trans-(4,5-dihydro-5-(3-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 09104

To a stirred and cooled (0° C.) solution of KOH (0.144 g, 2.57 mmol) inmethanol (10 mL) was added 3-iodobenzaldehyde (0.500 mg, 2.15 mmol) and1-isocyano-3-(pyrrolidin-1-yl)propan-2-one (0.295 g, 2.13 mmol). Thesolution was stirred 24 h with continued cooling and then concentrated.Water (50 mL) was added and the solution was extracted with EtOAc (3×50mL). The organic layer was washed with brine (50 mL), dried over MgSO₄,filtered and evaporated.trans-(4,5-Dihydro-5-(3-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 09104 was obtained (0.63 g, 80% yield) as a pale yellow solid.

MW: 370.19; Yield: 80%; Pale Yellow Solid.

¹H-NMR (CDCl₃, δ):1.80-2.08 (m, 4H, 2×CH₂), 3.42-3.58 (m, 3H, 1.5×CH₂),3.90-3.96 (m, 1H, 0.5×CH₂), 4.56 (dd, 1H, J=7.7 Hz, J=2.2 Hz, CH—N),6.10 (d, 1H, J=7.7 Hz, CH—O), 7.01 (d, 1H, J=2.2 Hz, CH═N), 7.11 (t, 1H,J=6.8 Hz, ArH), 7.29 (m, 1H, ArH), 7.66 (m, 2H, ArH).

¹³C-NMR (CDCl₃, δ): 24.2, 26.0, 46.5, 46.6, 75.8, 80.4, 94.7, 125.0,130.6, 134.6, 137.5, 142.0, 155.2, 166.3.

(±)-threo-2-Amino-3-hydroxy-3-(3-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 211

trans-(4,5-Dihydro-5-(3-iodophenyl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneSLA 09104 (0.620 g, 1.67 mmol) was dissolved in methanol (5 mL). Thesolution was stirred at room temperature and a solution of HCl (37%, 1mL) was added via syringe and the mixture was stirred at 50° C. for 6 h.The mixture was concentrated and triturated with EtOAc. After filtrationand drying(±)-threo-2-amino-3-hydroxy-3-(3-iodophenyl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 211 was obtained (586 mg, 88% yield) as a whitesolid.

MW: 396.65; Yield: 88%; White Solid; Mp (° C.): 183.7

¹H-NMR (CD₃OD, δ): 1.35-1.50 (m, 1H, 0.5×CH₂), 1.58-1.82 (m, 3H,1.5×CH₂), 2.08-2.18 (m, 1H, 0.5×CH₂), 3.21-3.45 (m, 4H, 2×CH₂), 4.09 (d,1H, J=9.1 Hz, CH—N), 4.80 (d, 1H, J=9.2 Hz, CH—O), 7.19 (t, 1H, J=7.1Hz, ArH), 7.49 (d, 1H, J=7.6 Hz, ArH), 7.74 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 360.9 ([MH]⁺, 100), 342.9 (40).

HPLC: Method A, detection UV 254 nm, Compound 211 RT=4.07 min, peak area93.4%.

Preparation of(±)-threo-2-amino-3-hydroxy-1-(4-methylpiperazin-1-yl)-3-(pyridin-4-yl)propan-1-onetrihydrochloride Compound 2122-Isocyano-1-(4-methyl-piperazin-1-yl)-ethanone VIB 01128

Prepared in accordance with Method B with methyl isocyanoacetate (1.0 g,10.09 mmol) and N-methylpiperazine (1.37 mL, 15.14 mmol). The reactionmixture was stirred overnight at RT and then concentrated. The residuewas dissolved in dichloromethane (50 mL) and evaporated. The residue wascoevaporated three times with a mixture of CH₂Cl₂:cyclohexane=50:50(3×10 mL). After drying 2-isocyano-1-(4-methyl-piperazin-1-yl)-ethanoneVIB 01128 was obtained as yellow solid (1.67 g, 99% yield).

MW: 167.21; Yield: 99%; Yellow Solid; Mp (° C.)=106.0

¹H NMR (CDCl₃, δ): 2.32 (m, 3H, Me) 2.38-2.50 (m, 4H, 2×CH₂), 3.42 (t,2H, J=4.7 Hz, CH₂), 3.66 (t, 2H, J=4.7 Hz, CH₂), 4.30 (s, 2H, CH₂).

¹³C-NMR (CDCl₃, δ): 42.4, 44.4, 45.5, 46.0, 54.3, 54.5, 160.8, 161.2.

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-methylpiperazin-1-yl)methanoneVIB 01130

To a stirred and cooled (0° C.) solution of KOH (0.335 g, 5.98 mmol) in7 mL MeOH were added successively pyridine-4-carbaldehyde (0.705 g, 6.58mmol) and 2-isocyano-1-(4-methyl-piperazin-1-yl)-ethanone VIB 01128(1.00 g, 6.58 mmol). The mixture was stirred at 0° C. untilprecipitation and concentrated. The mixture was partitioned betweenEtOAc (20 mL) and H₂O (10 mL). The aqueous layer was extracted twicewith EtOAc (60 mL). The EtOAc fractions were combined, washed twice withbrine (2×10 mL), dried over MgSO₄ and filtered. After evaporation anddryingtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-methylpiperazin-1-yl)methanoneVIB 01130 was obtained (1.282 g, 78% yield) as a yellow oil.

MW: 274.32; Yield: 78%; Yellow Oil.

¹H NMR (CDCl₃, δ): 2.32 (m, 3H, Me) 2.35-2.58 (m, 4H, 2×CH₂), 3.50-3.70(m, 2H, CH₂—N), 3.80-4.00 (m, 2H, CH₂—N), 4.59 (dd, 1H, J=7.8 Hz, J=2.2Hz, CH—N), 6.27 (d, 1H, J=7.8 Hz, O—CH), 7.02 (d, 1H, J=2.2 Hz, OCH═N),7.23 (d, 2H, J=4.7 Hz, ArH), 8.62 (dd, 2H, J=4.5 Hz, J=1.6 Hz, ArH).

¹³C-NMR (CDCl₃, δ): 42.6, 45.7, 46.0, 54.6, 55.1, 74.7, 79.6, 120.0(2×C), 148.6, 150.3 (2×C), 154.8, 166.0.

MS-ESI m/z (% rel. Int.): 275.2 ([MH]⁺, 40), 190.1 (35), 147.0 (40),127.0 (100).

HPLC: Method A, detection UV 254 nm, VIB 01130 RT=0.70 min, peak area99.9%.

(±)-threo-2-Amino-3-hydroxy-1-(4-methylpiperazin-1-yl)-3-(pyridin-4-yl)propan-1-onetrihydrochloride Compound 212

To a solution oftrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(4-methylpiperazin-1-yl)methanoneVIB 01130 (1.235 g, 4.50 mmol) in MeOH (15 mL) was added HCl 37% (1.6mL). After heating (50° C.) the mixture for 3.5 h a white solidprecipitated and the reaction mixture was concentrated and the crudeproduct was coevaporated twice with ethyl acetate. After triturationwith ethyl acetate, filtration and drying,(±)-threo-2-amino-3-hydroxy-1-(4-methylpiperazin-1-yl)-3-(pyridin-4-yl)propan-1-onetrihydrochloride Compound 212 (1.62 g, 96% yield) was obtained as awhite solid.

MW: 373.71; Yield: 96%; White Solid; Mp (° C.): 203.1

¹H-NMR (CD₃OD, δ): 2.95 (s, 3H, Me), 3.10-3.40 (m, 4H, 2×CH₂), 3.48-3.80(m, 2H, CH₂), 4.19-4.49 (m, 1H, 0.5×CH₂), 4.51-4.79 (m, 1H, 0.5×CH₂),5.06 (d, 1H, J=4.3 Hz, CH—NH₂), 5.35-5.63 (m, 1H, CH—O), 8.30 (s broad,2H, ArH), 8.95 (d, 2H, J=6.7 Hz, ArH).

MS-ESI m/z (% rel. Int.): 265.1 ([MH]⁺, 5), 248.1 (20), 156.1 (20),148.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 212 RT=0.68 min, peak area99.9%.

Preparation of(±)-threo-2-amino-1-(3,3-difluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 2131-(3,3-Difluoropyrrolidin-1-yl)-2-isocyanoethanone VIB 01158

To stirred and cooled (0° C.) methyl isocyanoacetate (96% technicalgrade, 345 mg, 3.48 mmol) was slowly added 3,3-difluoropyrrolidinehydrochloride (500 mg, 3.48 mmol), triethylamine (487 μL, 3.48 mmol) andMeOH (1 mL). The mixture was stirred for 15 h at RT and thenconcentrated. The resulting oil was coevaporated twice from EtOAc.1-(3,3-Difluoropyrrolidin-1-yl)-2-isocyanoethanone VIB 01158 wasobtained as a yellow oil (305 mg, 60% yield) and used in the next stepwithout purification.

MW: 174.17; Yield: 60%; Yellow Oil.

¹H NMR (CDCl₃, δ): 2.30-2.62 (m, 2H, CH₂) 3.65-3.90 (m, 4H, CH₂), 4.25(d, 2H, J=17.5 Hz, CH₂).

trans-(3,3-Difluoropyrrolidin-1-yl)(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanoneVIB 01160

To a stirred and cooled (0° C.) solution of KOH (0.098 g, 1.75 mmol) inMeOH (2 mL) were added successively pyridine-4-carbaldehyde (0.206 g1.92 mmol) and 1-(3,3-difluoropyrrolidin-1-yl)-2-isocyanoethanone VIB01158 (0.305 g, 1.75 mmol). The mixture was stirred at 0° C. for 3 h.After evaporation of MeOH, the mixture was partitioned between EtOAc (50mL) and H₂O (40 mL). The aqueous layer was extracted with EtOAc (4×50mL). The fractions were combined, washed twice with brine (2×20 mL),dried over MgSO₄ and filtered. After evaporation and dryingtrans-(3,3-difluoropyrrolidin-1-yl)(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanoneVIB 01160 (0.33 g, 67% yield) was obtained as a yellow oil.

MW: 281.26; Yield: 67%; Yellow Oil.

¹H NMR (CDCl₃, δ): 2.30-2.60 (m, 2H, CH₂) 3.67-3.99 (m, 3H, 1.5×CH₂),4.22-4.59 (m, 2H, CH—N & 0.5×CH₂), 6.17 (d, 1H, J=7.8 Hz, CH—O), 7.04(dd, 1H, J=3.2 Hz, J=2.5 Hz, HC═N), 7.20-7.38 (m, 2H, ArH), 8.55-8.70(m, 2H, ArH).

(±)-threo-2-Amino-1-(3,3-difluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 213

To a solution oftrans-(3,3-difluoropyrrolidin-1-yl)(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanoneVIB 01160 (0.305 g, 1.08 mmol) in methanol (4 mL) was added hydrochloricacid 37% (395 μL). After heating (50° C.) the mixture for 3.5 h thereaction mixture was concentrated and the crude product was coevaporatedtwice with EtOAc. After trituration with EtOAc, filtration and drying,(±)-threo-2-amino-1-(3,3-difluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 213 (269 mg, 44% yield) was obtained as a beigesolid.

MW: 344.19; Yield: 44%; Beige Solid; Mp (° C.): 182.2

¹H-NMR (CD₃OD, δ): 2.25-2.62 (m, 2H, CH₂), 3.30-4.20 (m, 4H, CH₂), 4.55(d, 1H, J=5.5 Hz, 0.35×CH—N), 4.66 (d, 1H, J=5.5 Hz, 0.65×CH—N), 5.42(m, 1H, CH—O), 8.23 (d, 2H, J=4.8 Hz, ArH), 8.94 (d, 2H, J=5.9 Hz, ArH).

MS-ESI m/z (% rel. Int.): 272.2 ([MH]⁺, 15), 254.1 (15), 178.1 (20),148.1 (100), 137.1 (95).

HPLC: gradient of solvent B:solvent A=5:95 to 100% solvent B in 7 min.Solvent A was H₂O with 0.1% Et₃N and solvent B was CH₃CN with 0.1% Et₃N;detection UV 254 nm, Compound 213 RT=4.70 min, peak area 98.1%.

Preparation of (2S,3R)- &(2R,3S)-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compounds 2141-((S)-3-Fluoropyrrolidin-1-yl)-2-isocyanoethanone VIB 01166

To stirred and cooled (0° C.) methyl isocyanoacetate (96% technicalgrade, 1.18 g, 11.9 mmol) was slowly added (S)-(+)-3-fluoropyrrolidinehydrochloride (97%, 1.5 g, 11.9 mmol), triethylamine (1.67 mL, 11.9mmol) and MeOH (3 mL). The mixture was stirred for 15 h at RT andconcentrated. The mixture was stirred for 15 h at RT and concentrated.Water was added (50 mL) and the mixture was extracted with EtOAc (3×50mL), dried over MgSO₄, filtered and evaporated to obtained crude1-((S)-3-fluoropyrrolidin-1-yl)-2-isocyanoethanone VIB 01166 (1.47 g,79% yield) as a brown oil which was used in the next step withoutpurification.

MW: 156.152; Yield: 79%; Brown Oil.

¹H NMR (CDCl₃, δ): 1.85-2.50 (m, 2H, CH₂), 3.40-4.35 (m, 6H, 3×CH₂),5.17-5.47 (m, 1H, CHF).

MS-ESI m/z (% rel. Int.): 171.1 ([MH⁺+Na], 100), 157.1 ([MH]⁺, 82),130.1 (95).

trans-((S)-3-Fluoropyrrolidin-1-yl)((4S,5R)- &4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01168

To a stirred and cooled (0° C.) solution of KOH (0.526 g, 9.39 mmol) inMeOH (10 mL) were added successively pyridine-4-carbaldehyde (1.10 g,10.33 mmol) and 1-((S)-3-fluoropyrrolidin-1-yl)-2-isocyanoethanone VIB01166 (1.47 g, 9.39 mmol). The mixture was stirred at 0° C. to RT for 24h. After evaporation of MeOH, the mixture was partitioned between EtOAc(40 mL) and H₂O (20 mL). The aqueous layer was extracted with EtOAc(6×40 mL). The EtOAc fractions were combined, washed twice with brine(2×10 mL), dried over MgSO₄, filtered and evaporated. The crude productwas purified by column chromatography (florisil, EtOAc:MeOH=9:1). Afterevaporation and drying trans-((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- &4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01168 (590mg, diastereoisomeric mixture in ratio 1:1, 24% yield) were obtained asa yellow oil.

MW: 263.27; Yield: 24%; Yellow Oil.

¹H NMR (CDCl₃, δ): 1.89-2.46 (m, 2H, CH₂) 3.5-4.45 (m, 4H, CH₂),4.46-4.60 (m, 1H, CH—N), 5.16-5.45 (m, 1H, CH—F), 6.12-6.25 (m, 1H,CH—O), 6.95-7.18 (m, 1H, CH═N), 7.20-7.40 (m, 2H, ArH), 8.50-8.70 (m,2H, ArH).

(2S,3R)- &(2R,3S)-2-Amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compounds 214

To a solution of ((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- &4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01168(0.590 g, 2.24 mmol) in methanol (8 mL) was added hydrochloric acid 37%(686 μL). After heating (50° C.) the mixture for 3.5 h the reactionmixture was concentrated and the crude product was coevaporated twicewith EtOAc. After trituration with EtOAc, filtration and drying (2S,3R)-&(2R,3S)-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compounds 214 (474 mg, diastereoisomeric mixture inratio 1:1, 68% yield) were obtained as a pale yellow solid.

MW: 326.2; Yield: 68%; Pale Yellow Solid; Mp (° C.): 173.1.

¹H-NMR (CD₃OD, δ): 1.82-2.38 (m, 2H, CH₂), 2.80-4.15 (m, 4H, 2×CH₂),4.35-4.68 (m, 1H, CH—N), 5.00-5.50 (m, 2H, CH—O & CH—F), 8.11-8.32 (m,2H, ArH), 8.82-9.00 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 254.2 ([MH]⁺, 15), 237.1 (20), 148.1 (100),137.1 (70).

HPLC: gradient of solvent B:solvent A=5:95 to 100% solvent B in 7 min.Solvent A was H₂O with 0.1% Et₃N and solvent B was CH₃CN with 0.1% Et₃N;detection UV 254 nm, Compounds 214 RT=4.35 min, peak area 99.0%.

Preparation of (2S,3R)- &(2R,3S)-2-amino-1-((R)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compounds 2151-((R)-3-Fluoropyrrolidin-1-yl)-2-isocyanoethanone BLE 04170

To stirred and cooled (0° C.) methyl isocyanoacetate (96% technicalgrade, 0.79 g, 7.96 mmol) was slowly added (R)-(−)-3-fluoropyrrolidinehydrochloride (1.0 g, 7.96 mmol), triethylamine (1.11 mL, 7.96 mmol) andMeOH (2.5 mL). The mixture was stirred for 15 h at RT and concentrated.Water was added (50 mL) and the mixture was extracted with EtOAc (3×50mL), dried over MgSO₄, filtered and evaporated to obtained crude1-((R)-3-fluoropyrrolidin-1-yl)-2-isocyanoethanone BLE 04170 as a brownoil (0.96 g, 77% yield) which was used in the next step without furtherpurification.

MW: 156.152; Yield: 77%; Brown Oil.

¹H NMR (CDCl₃, δ): 1.85-2.50 (m, 2H, CH₂), 3.40-4.35 (m, 6H, 3×CH₂),5.17-5.47 (m, 1H, CHF).

trans-((R)-3-Fluoropyrrolidin-1-yl)((4S,5R)- &4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones BLE 04172

To a stirred and cooled (0° C.) solution of KOH (0.34 g, 6.06 mmol) inMeOH (4.5 mL) were added successively pyridine-4-carbaldehyde (0.71 g,6.66 mmol) and a solution of BLE 04170 (0.95 g, 6.06 mmol) in MeOH (2.5mL). The mixture was stirred at 0° C. to RT for 15 h. After evaporationof MeOH, the mixture was partitioned between EtOAc (40 mL) and H₂O (20mL). The aqueous layer was extracted with EtOAc (3×40 mL). The EtOAcfractions were combined, washed with brine (50 mL), dried over MgSO₄,filtered and evaporated. The crude product was purified by columnchromatography (florisil, EtOAc:MeOH=9:1). After evaporation and dryingtrans-((R)-3-fluoropyrrolidin-1-yl)((4S,5R)- &4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones BLE 04172 (653mg, diastereoisomeric mixture in ratio 1:1, 41% yield) were obtained asa colorless oil.

MW: 263.27; Yield: 41%; Colorless Oil.

¹H NMR (CDCl₃, δ): 1.89-2.46 (m, 2H, CH₂) 3.5-4.45 (m, 4H, 2×CH₂),4.46-4.60 (m, 1H, CH—N), 5.16-5.45 (m, 1H, CH—F), 6.12-6.25 (m, 1H,CH—O), 6.95-7.18 (m, 1H, CH═N), 7.20-7.40 (m, 2H, ArH), 8.50-8.70 (m,2H, ArH).

(2S,3R)- &(2R,3S)-2-Amino-1-((R)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compounds 215

To a solution of trans-((R)-3-fluoropyrrolidin-1-yl)((4S,5R)- &4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones BLE 04172(0.65 g, 2.47 mmol) in methanol (8 mL) was added hydrochloric acid 37%(757 μL). After heating (50° C.) the mixture for 3.5 h the reactionmixture was concentrated and the crude product was coevaporated twicewith EtOAc. After trituration with EtOAc, filtration and drying (2S,3R)-&(2R,3S)-2-amino-1-((R)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compounds 215 (544 mg, diastereoisomeric mixture inratio 1:1, 68% yield) were obtained as a pale yellow solid.

MW: 326.2; Yield: 68%; Pale Yellow Solid; Mp (° C.): 134.5

¹H-NMR (CD₃OD, δ): 1.82-2.38 (m, 2H, CH₂), 2.80-4.15 (m, 4H, CH₂),4.35-4.68 (m, 1H, CH—N), 5.00-5.50 (m, 2H, CH—O & CHF), 8.11-8.32 (m,2H, ArH), 8.82-9.00 (m, 2H, ArH).

Preparation ofN-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)formamidehydrochloride Compound 216

trans-(4,5-Dihydro-5-(pyridin-4-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneCompound 19 (0.200 g, 0.81 mmol) was dissolved in methanol (5 mL). Dowex50WX8-200 (0.5 mL, washed beforehand by a 0.5 M solution of HCl thenwater) was added at RT. The mixture was stirred for 1 h at 50° C. andafter cooling was filtered off. MeOH was evaporated and the residue wasdried to give 150 mg of a pasty product (150 mg, 61.0% yield). The freebase form was dissolved in a minimum of a mixture of EtOAc:MeOH=95:5 anda 0.4 M solution of HCl in ether (1.3 mL, 0.52 mmol) was added at +4° C.After evaporation of solvents, the product was precipitated with diethylether, filtered and dried to obtainN-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)formamidehydrochloride Compound 216 as a beige solid (100 mg, 41.0% yield).

MW: 299.75; Yield: 41.0%; Beige Solid; Mp (° C.): 203.1

R_(f): 0.35 (CH₂Cl₂:MeOH=90:10) free base.

¹H-NMR (CD₃OD, *): 1.87-2.00 (m, 4H, 2×CH₂), 3.41-3.46 (m, 2H, CH₂—N),3.62-3.66 (m, 2H, CH₂—N), 5.18 (d, 1H, J=3.6 Hz, CH—N), 5.40 (d, 1H,J=3.8 Hz, CH—O), 7.99 (s, 1H, CH═O), 8.16 (d, 2H, J=5.8 Hz, ArH), 8.81(d, 2H, J=5.5 Hz, ArH).

¹³C-NMR (CD₃OD, *): 25.0, 27.0, 47.5, 48.3, 55.6, 72.4, 126.6 (2×C),142.1 (2×C), 163.4, 164.1, 168.5.

MS-ESI m/z (% rel. Int.): 264.1 ([MH]⁺, 10), 148.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 216 RT=1.30 min, peak area98.0%.

Preparation ofN-((±)-threo-2-acetoxy-2-pyridin-4-yl-1-(pyrrolidine-1-carbonyl)-ethyl)-formamidehydrochloride Compound 217

N-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)formamidehydrochloride Compound 216 (0.220 g, 0.80 mmol) was dissolved in CH₂Cl₂(10 mL) with triethylamine (280 μL, 2 mmol) at 0° C. Then aceticanhydride (160 μL, 1.6 mmol) was added slowly and the mixture wasstirred for 72 h at RT. The solvent was evaporated and the residue wasdried under vacuum. After column chromatography (SiO₂, EtOAc:MeOH=90:10)to give a pasty product (100 mg, 41% yield). The product obtained wasdissolved in MeOH (10 mL) and a solution of HCl (0.4 M, 1 mL) in Et₂Owas added at 0° C. Evaporation of the volatiles led to acetic acidN-((±)-threo-2-acetoxy-2-pyridin-4-yl-1-(pyrrolidine-1-carbonyl)-ethyl)-formamidehydrochloride Compound 217 was obtained as a white solid (110 mg, 40%yield).

MW: 341.79; Yield: 40.0%; White Solid; Mp (° C.): 173.9.

R_(f): 0.25 (EtOAc:MeOH=90:10, free base).

¹H-NMR (CD₃OD,*): 1.80-2.00 (m, 4H, 2×CH₂), 2.19 (s, 3H, CH₃) 3.29-3.64(m, 4H, 2×CH₂—N), 5.34 (s, 1H, N—CH), 6.44 (s, 1H, O—CH), 8.01 (s, 1H,CHO), 8.16 (s, 2H, ArH), 8.85 (s, 2H, ArH).

¹³C-NMR (CD₃OD, *): 20.5, 25.0, 27.0, 47.6, 47.7, 53.9, 73.4, 126.7(2×C), 142.8 (2×C), 144.3, 163.3, 166.9, 170.9.

MS-ESI m/z (% rel. Int.): 306.1 ([MH]⁺, 10), 261.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 217 RT=0.90 min, peak area97.0%.

Preparation of tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamatehydrochloride Compound 218

(±)-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (2.90 g, 9.3 mmol) was dissolved in CH₂Cl₂(250 mL) with Et₃N (4.3 mL, 30.7 mmol) at 0° C. Di-tert-butyldicarbonate (2.45 g, 11.2 mmol) in CH₂Cl₂ (50 mL) was added slowly andthe mixture was stirred for 15 h at RT. Brine (50 mL) was added and theproduct was extracted with CH₂Cl₂. After drying over MgSO₄ andfiltration, CH₂Cl₂ was evaporated and the residue was dried in vacuum.After column chromatography (SiO₂, EtOAc:MeOH=90:10), tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08100 was obtained as a beige solid (2.00 g, 64% yield). A Sample ofTTA 08100 (55 mg) was dissolved in CH₂Cl₂ (1 mL) and Et₂O (30 mL) and asolution of HCl (0.1 M, 2 mL) in Et₂O was added at 0° C. Evaporation ofthe volatiles led to tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamatehydrochloride Compound 218 (50 mg, 52% yield) as a white solid.

MW: 371.86; Yield: 52.0%; White Solid; Mp (° C.): 141.2.

R_(f): 0.30 (EtOAc:MeOH=90:10, free base).

¹H-NMR (CD₃OD, *): 1.33 (s, 9H, 3×CH₃) 1.91-2.00 (m, 4H, 2×CH₂),3.44-3.49 (m, 2H, CH₂), 3.60-3.64 (m, 2H, CH₂—N), 4.78 (s, 1H, N—CH),5.38 (s, 1H, O—CH), 8.16 (s, 2H, ArH), 8.83 (s, 2H, ArH).

¹³C-NMR (CD₃OD, *): 25.0, 27.1, 28.5 (3×C), 47.5, 48.0, 58.3, 72.8,81.0, 126.7 (2×C), 142.0 (2×C), 157.3, 164.6, 169.5.

MS-ESI m/z (% rel. Int.): 336.1 ([MH]⁺, 20), 219.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 218 RT=3.8 min, peak area98.0%.

Preparation of(±)-erythro-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 219 tert-Butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamateVIB 01080

To a solution of(±)-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (3.25 g, 10.54 mmol) in CH₂Cl₂ (250 mL) in a500 mL round bottom flask equipped with a magnetic stirrer and undernitrogen atmosphere was added via syringe at 0° C. triethylamine (4.69mL, 33.73 mmol). A solution of di-tert-butyldicarbonate (2.76 g, 12.65mmol) in CH₂Cl₂ (75 mL) was added at 0° C. dropwise via a droppingfunnel. The reaction mixture was abandoned at 0° C. for 2 h then at RTovernight. A solution of brine (130 mL) was added and the solution wasextracted with CH₂Cl₂ (3×75 mL), dried over MgSO₄, filtered andevaporated. The crude product was purified by column chromatography(SiO₂, EtOAc:MeOH=90:10). After evaporation to dryness of the combinedfractions, tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamateVIB 01080 (2.79 g, 79% yield) was obtained as a yellow solid.

MW: 335.4; Yield: 79%; Yellow Solid; Mp (° C.): 160.5

R_(f): 0.31 (EtOAc:MeOH=90:10).

¹H-NMR (CDCl₃, *: 1.26 (s, 9H, 3×CH₃), 1.71-2.00 (m, 4H, 2×CH₂),3.25-3.60 (m, 4H, 2×CH₂N), 4.61 (dd, 1H, J=9.8 Hz, J=2.6 Hz, N—CH), 4.96(s, 1H, OH), 5.08 (d, 1H, J=2.6 Hz, NH), 5.47 (d, 1H, J=9.8 Hz, O—CH),7.35 (d, 2H, J=5.7 Hz, ArH), 8.58 (d, 2H, J=5.4 Hz, ArH).

¹³C-NMR (CDCl₃, *): 24.0 (2×C), 25.9 (2×C), 28.1 (3×C), 46.1, 46.7,56.1, 60.4, 72.8, 80.2, 121.4 (2×C), 148.7, 149.5 (2×C), 155.6, 169.4.

MS-ESI m/z (% rel. Int.): 336.1 ([MH]⁺, 45), 280 (18), 219 (100); 148(38).

tert-Butyl(±)-1,3-dioxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08120

To a solution of tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamateVIB 01080 (3.92 g, 11.70 mmol) in CH₂Cl₂ (320 mL) in a 500 mL roundbottom flask equipped with a magnetic stirrer and under nitrogenatmosphere was added slowly Dess-Martin periodinane (4.96 g, 11.70 mmol)at RT. The reaction mixture was stirred at RT for 0.5 h and CH₂Cl₂,washed with a mixture of saturated sodium bicarbonate (100 mL), 1 Msodium thiosulfate (50 mL), brine (50 mL) and dried over MgSO₄, filteredand evaporated. Diethyl ether (250 mL) was added and the precipitate wasfiltered off. After evaporation of the filtrate, the crude product waspurified by column chromatography (SiO₂, CH₂Cl₂:EtOAc=4:6). Afterevaporation to dryness of the combined fractions tert-butyl(±)-1,3-dioxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08120 (3.0 g, 77% yield) was obtained as a white solid.

MW: 333.38; Yield: 77%; White Solid; Mp (° C.): 125.4

R_(f): 0.25 (CH₂Cl₂:EtOAc=4:6).

¹H-NMR (CDCl₃, *): 1.36 (s, 9H, 3×CH₃), 1.86-2.00 (m, 4H, 2×CH₂),3.47-3.71 (m, 4H, 2×CH₂N), 5.62 (d, 1H, J=7.4 Hz, N—CH), 6.07 (d, 1H,J=7.4 Hz, NH), 7.80 (dd, 2H, J=4.6 Hz, J=1.3 Hz, ArH), 8.80 (dd, 2H,J=4.5 Hz, J=1.5 Hz, ArH).

¹³C-NMR (CDCl₃, *): 24.0, 26.0, 28.1 (3×C), 46.8, 47.0, 61.0, 80.9,121.5 (2×C), 141.4, 150.8 (2×C), 155.1, 163.8, 194.3.

MS-ESI m/z (% rel. Int.): 334.1 ([MH]⁺, 10), 173.1 (30), 129.1 (100).

tert-Butyl(±)-erythro-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08124P

To a solution of tert-butyl(±)-1,3-dioxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08120 (2.28 g, 6.80 mmol) in MeOH (50 mL) in a 250 mL round bottomflask equipped with a magnetic stirrer and under nitrogen atmosphere wasadded slowly sodium borohydride (285 mg, 7.50 mmol) at RT. The reactionmixture was stirred at RT for 0.5 h and was cooled at 4° C. A solutionof 2 M NaOH (10 mL) was added and MeOH was evaporated at 30° C. EtOAc(300 mL) was added and the organic phase was washed with brine (20 mL),dried over MgSO₄, filtered and evaporated to give a crude compound(ratio erythro:threo=80:20 estimated by ¹H NMR 300 MHz) TTA 08124 (2.1g, 92% yield). The crude product was recrystallized in EtOAc and afterfiltration and drying tert-butyl(±)-erythro-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08124P (1.30 g, ratio erythro:threo=96:4 estimated by ¹H NMR 300 MHzanalysis, 57% yield) was obtained as a white solid.

MW: 335.40; Yield: 57%; White Solid; Mp (° C.): 170.3

R_(f): 0.45 (EtOAc: MeOH=85:15).

¹H-NMR (CDCl₃, *): 1.43 (s, 9H, 3×CH₃), 1.64-1.83 (m, 4H, 2×CH₂),2.88-2.96 (m, 1H, CH₂N), 3.23-3.29 (m, 1H, CH₂N), 3.34-3.43 (m, 1H,CH₂N), 3.56-3.63 (m, 1H, CH₂N), 4.66 (dd, 1H, J=8.9 Hz, J=3.7 Hz, N—CH),4.91 (dd, 1H, J=8.4 Hz, J=3.4 Hz, O—CH), 5.42 (d, 1H, J=8.8 Hz, OH),5.75 (d, 1H, J=8.8 Hz, NH), 7.30 (d, 2H, J=5.9 Hz, ArH), 8.57 (d, 2H,J=5.8 Hz, ArH). Ratio erythro:threo=96:4.

¹³C-NMR (CDCl₃, *): 24.0, 25.7, 28.1 (3×C), 45.7, 46.8, 55.2, 74.5,80.5, 120.9 (2×C), 149.6, 149.8 (2×C), 155.4, 168.8.

MS-ESI m/z (% rel. Int.): 336.1 ([MH]⁺, 10), 280.1 (20), 110.0 (100).

(±)-erythro-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 219

To a solution of tert-butyl(±)-erythro-3-hydroxy-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08124P (1.15 g, 3.40 mmol) in MeOH (30 mL) in a 250 mL round bottomflask equipped with a magnetic stirrer was added HCl 37% (3 mL, 35 mmol)at RT. The reaction mixture was stirred at RT for 0.4 h at 45° C. andMeOH was evaporated at 45° C. to give after drying, a white solid TTA08136 (ratio erythro:threo=96:4 estimated by ¹H NMR 300 MHz analysis).Amberlite IRA-400 (Cl⁻) (10 g) was washed successively with water (2×10mL), 0.5 N NaOH (3×20 mL), water (2×10 mL) and MeOH (3×10 mL). Asolution of TTA 08136 in MeOH (30 mL) was stirred with washed AmberliteIRA-400 for 5 min at RT. After filtration, the MeOH was evaporated andthe free base form was purified by column chromatography (SiO₂,CHCl₃:EtOH 95°=86:14) to give TTA 08136A (445 mg, 55% yield) as a beigesolid (no threo isomer detected by ¹H NMR 300 MHz and HPLC). TTA 08136A(193 mg) was stirred at RT in ethyl acetate (5 mL) with a solution of0.1 N HCl in isopropanol (17 mL). Solvents were evaporated at 33° C.,MeOH (0.5 mL) was added and the salt was precipitated by addition ofEtOAc (20 mL) to give nearly quantitatively after filtration and drying(±)-erythro-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 219 as a white solid (no threo isomer detectedby ¹H NMR 300 MHz and HPLC).

MW: 308.20; Yield: 55%; White Solid; Mp (° C.): 154.1.

R_(f): 0.18 (CHCl₃:EtOH 95°=86:14, free base).

¹H-NMR (CD₃OD, *): 1.94-2.04 (m, 4H, 2×CH₂), 3.45-3.56 (m, 2H, CH₂N),3.66-3.78 (m, 2H, CH₂N), 4.71 (d, 1H, J=5.2 Hz N—CH), 5.50 (d, 1H, J=5.1Hz, O—CH), 8.12 (d, 2H, J=5.6 Hz, ArH), 8.92 (d, 2H, J=5.4 Hz, ArH).

¹³C-NMR (CD₃OD, *): 25.0, 27.0, 47.8, 58.4, 70.0, 127.2 (2×C), 143.1(2×C), 159.8, 164.8.

MS-ESI m/z (% rel. Int.): 236.1 ([MH]⁺, 10), 219.1 (55), 110.0 (100).

Preparation of(−)-(2R,3R)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 220 and(+)-(2S,3S)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 221 Analytical Chiral Separation:

20 μL of a 1 mg/mL solution of Compound 219 were injected on ChiralpakAD: flow-rate=1 mL/min, temperature=25° C., mobile phase:hexane:ethanol=8:2, detection UV at 220 nm and by polarimeter, Rt(−)=16.26 min, Rt (+=19.02 min, k(−)=4.38, k(+)=5.30, α=1.21 andresolution Rs=1.90.

Semi-Preparative Separation was Performed on Chiralpak AS (250×10 mm):

A 40 mg/mL solution of(±)-erythro-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneCompound 219 racemate was prepared and 100 μL of this solution wereinjected every 4 min on Chiralpak AS, flow-rate=5 mL/min, mobile phasehexane:ethanol=1:1, UV detection at 254 nm. 56 successive injectionswere done. The two main fractions were identified on UW and collected intwo different flasks. The solvent was removed in vacuo at 30° C. Theresulting solid was dissolved in 50 mL of CH₂Cl₂ and then filtered on a0.45 μm millipore membrane. After evaporation of CH₂Cl₂, the solid wasdissolved in 50 mL of methanol and then filtered. The salt wasregenerated according to the procedure reported for Compound 203 andCompound 204.

The enantiomeric purity of the compounds was checked by analyticalinjection on Chiralpak AD of the regenerated salts:

(−)-(2R,3R)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 220

MW: 308.20; 133 mg obtained; White Solid; Mp (° C.): too hygroscopic.

R_(f): 0.18 (CHCl₃:EtOH 95°=86:14, free base).

Enantiomeric excess=99.1% measured by HPLC at 220 nm (Chiralpak AD)

α²⁵ _(D)=−6.4 (MeOH, c=1)

¹H-NMR (CD₃OD, δ): 1.94-2.03 (m, 4H, 2×CH₂), 3.45-3.55 (m, 2H, CH₂—N),3.63-3.76 (m, 2H, CH₂—N), 4.68 (d, 1H, J=5.1 Hz, N—CH), 5.48 (d, 1H,J=5.5 Hz, O—CH), 8.08 (d, 2H, J=6.4 Hz, ArH), 8.90 (d, 2H, J=6.6 Hz,ArH).

¹³C-NMR (CD₃OD, δ): 23.4, 25.4, 46.2, 56.7, 68.4, 125.7 (2×C), 141.3(2×C), 158.5, 163.2.

(+)-(2S,3S)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 221

MW: 308.20; 140 mg obtained; White Solid; Mp (° C.): too hygroscopic.

R_(f): 0.18 (CHCl₃:EtOH 95°=86:14, free base).

Enantiomeric excess=99.1% measured by HPLC at 220 nm (Chiralpak AD)

α²⁵ _(D)=+6.3 (MeOH, c=1).

¹H-NMR (CD₃OD, δ): 1.94-2.02 (m, 4H, 2×CH₂), 3.44-3.52 (m, 2H, CH₂—N),3.64-3.74 (m, 2H, CH₂—N), 4.68 (d, 1H, J=5.2 Hz, N—CH), 5.48 (d, 1H,J=4.3 Hz, O—CH), 8.09 (d, 2H, J=6.4 Hz, ArH), 8.90 (d, 2H, J=6.1 Hz,ArH).

¹³C-NMR (CD₃OD, δ): 23.4, 25.4, 46.2, 56.8, 68.4, 125.7 (2×C), 141.3(2×C), 158.5, 163.2.

Preparation of(±)-threo-3-hydroxy-2-(isopropylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 223

To a stirred solution of(±)-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (0.20 g, 0.65 mmol) in 3 mL of MeOH wereadded triethylamine (180 μl, 1.30 mmol) and acetone (75 μL, 1.00 mmol).The mixture was stirred overnight at RT under nitrogen and then wasadded AcOH (200 μL, 3.2 mmol) and NaBH₃CN (85 mg, 1.3 mmol). After 5 hat 20° C., MeOH was evaporated and the residue was partitioned betweenCH₂Cl₂ and 1 N aqueous sodium carbonate. The organic layer wasevaporated and the obtained residue was purified by columnchromatography (SiO₂, EtOAc:MeOH=9:1). HCl treatment in MeOH gave(±)-threo-3-hydroxy-2-(isopropylamino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 223 (56 mg, 25% yield) as a white solid.

MW: 350.28; Yield: 25%; White Solid; Mp (° C.): 189.8.

R_(f): 0.20 (EtOAc:MeOH=9:1, free base).

¹H-NMR (CD₃OD, δ): 1.32 (d, 3H, J=6.5 Hz, CH₃), 1.40 (d, 3H, J=6.5 Hz,CH₃), 1.55-1.84 (m, 4H, 2×CH₂), 2.66-2.74 (m, 1H, CH), 3.27-3.54 (m, 4H,CH₂—N), 4.60 (d, 1H, J=7.9 Hz, N—CH), 5.38 (d, 1H, J=7.8 Hz, O—CH), 8.24(d, 2H, J=6.3 Hz, ArH), 8.98 (d, 2H, J=6.3 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 18.7, 20.1, 24.7, 26.7, 47.7, 48.5, 52.2, 62.4,72.5, 127.0 (2×C), 143.2 (2×C), 161.1, 164.1.

MS-ESI m/z (% rel. Int.): 278.1 ([MH]⁺, 25), 179.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 223 RT=1.30 min, peak area99.0%.

Preparation of(±)-2-amino-3-(hydroxyimino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 224(±)-2-tert-Butyloxycarbonylamino-3-(hydroxyimino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneTTA 08160

To a stirred solution of tert-butyl(±)-1,3-dioxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamateTTA 08120 (0.20 g, 0.60 mmol) in 10 mL of dioxane were added Et₃N (125μl, 0.90 mmol) and hydroxylamine hydrochloride (65 mg, 0.90 mmol). Themixture was stirred 2 h at 110° C. in a sealed tube then dioxane wasevaporated. The obtained residue was purified by column chromatography(SiO₂, CH₂Cl₂:MeOH=9:1 to 97:3) to give(±)-2-tert-butyloxycarbonylamino-3-(hydroxyimino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneTTA 08160 (100 mg, 48% yield) as an oil.

MW: 348.40; Yield: 48%; Oil.

R_(f): 0.30 (CH₂Cl₂:MeOH=97:3).

¹H-NMR (CDCl₃, δ): 1.39 (s, 9H, 3×CH₃), 1.81-1.98 (m, 4H, 2×CH₂),3.37-3.62 (m, 4H, 2×CH₂—N), 5.44 (d, 1H, J=8.1 Hz, N—CH), 5.97 (d, 1H,J=8.1 Hz, NH), 7.37 (d, 2H, J=5.6 Hz, ArH), 8.65 (d, 2H, J=6.0 Hz, ArH).

¹³C-NMR (CDCl₃, δ): 24.0, 26.0, 28.2 (3×C), 46.5, 46.7, 55.8, 80.3,123.1 (2×C), 139.7, 149.4 (2×C), 155.1, 166.4.

MS-ESI m/z (% rel. Int.): 349.2 ([MH]⁺, 85), 293.2 (100).

HPLC: Method A, detection UV 254 nm, TTA 08160 RT=3.90 min, peak area97.0%.

(±)-2-Amino-3-(hydroxyimino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 224

(±)-2-tert-Butyloxycarbonylamino-3-(hydroxyimino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneTTA 08160 (100 mg, 0.29 mmol) was dissolved in MeOH (2 mL) and asolution of 1 M HCl in MeOH (1 mL, 3.00 mmol) was added and the mixturewas heated for 10 min at 45° C. MeOH was evaporated and the residue wasdried to give crude(±)-2-amino-3-(hydroxyimino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneTTA 08164. Amberlite IRA-400 (Cl⁻) (1 mL, 1.4 mmol) was washedsuccessively with water (2×10 mL), NaOH 0.5 N (3×20 mL), water (2×10 mL)and MeOH (3×10 mL). A solution of TTA 08164 in MeOH (30 mL) was stirredwith washed Amberlite IRA-400 for 5 min at RT. After filtration, theMeOH was evaporated and the free base form was purified by columnchromatography (SiO₂, CH₂Cl₂:MeOH=9:1). HCl Treatment in MeOH gave(±)-2-amino-3-(hydroxyimino)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 224 (29 mg, 31% yield) as a beige solid.

MW: 321.20; Yield: 31%; Beige Solid; Mp (° C.): 225.2

R_(f): 0.30 (EtOAc:MeOH=9:1, free base).

¹H-NMR (CD₃OD, δ): 1.85-2.04 (m, 4H, 2×CH₂), 3.23-3.73 (m, 4H, CH₂—N),5.58 (d, 1H, J=4.4 Hz, CH), 8.23 (d, 2H, J=5.0 Hz, ArH), 9.03 (d, 2H,J=5.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.9, 47.9, 48.0, 55.6, 128.7 (2×C), 143.8(2×C), 145.5, 148.8, 163.8.

MS-ESI m/z (% rel. Int.): 249.2 ([MH]⁺, 10), 115.0 (100).

HPLC: Method A, detection UV 254 nm, Compound 224 RT=0.60 min, peak area99.0%.

Preparation of(±)-threo-2-(4-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 225

To a stirred solution of(±)-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (500 mg, 1.62 mmol) in 7.5 mL of MeOH at RTunder nitrogen was added dropwise triethylamine (495 μL, 3.56 mmol) and4-iodobenzaldehyde (413 mg, 1.73 mmol). The mixture was stirred for 5 hat RT under nitrogen. Acetic acid (463 μL, 8.1 mmol) and NaBH₃CN (356mg, 5.67 mmol) were added. The mixture was stirred for another 15 h atRT. The mixture was partitioned between EtOAc (750 mL) and an 10%aqueous solution of potassium carbonate. The organic layer was washedwith brine (2×20 mL), dried over MgSO₄ and filtered. After evaporation,the crude product was purified by column chromatography (SiO₂,EtOAc:MeOH=85:15) to give(±)-threo-2-(4-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneVIB 01096 as a yellow solid (380 mg, 52% yield). To a solution of(±)-threo-2-(4-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-oneVIB 01096 (380 mg, 8.43 mmol) in methanol (10 mL) was added a solutionof 0.5 M aqueous hydrochloric acid (7 mL). After stirring the mixture atRT for 0.5 h the reaction mixture was concentrated and the crude productwas coevaporated twice with EtOAc. After trituration with EtOAc,filtration and drying,(±)-threo-2-(4-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onehydrochloride Compound 225 (373 mg, 44% yield) was obtained as a paleyellow solid.

MW: 524.37; Yield 44%; Pale Yellow Solid; Mp (° C.): 194.7

¹H-NMR (CD₃OD, δ): 1.42-1.80 (m, 4H, 2×CH₂), 2.32-2.50 (m, 1H, CH₂),2.96-3.12 (m, 1H, CH₂), 3.12-3.25 (m, 2H, CH₂), 4.27 (q, 2H, J=13.3 Hz,CH₂), 4.42 (d, 1H, J=7.8 Hz, HC—N), 5.31 (d, 2H, J=7.7 Hz, HC—O), 7.29(d, 2H, J=8.1 Hz, ArH), 7.80 (dd, 2H, J=2.9 Hz, J=8.1 Hz, ArH), 8.15 (d,2H, J=5.5 Hz, ArH), 8.91 (d, 2H, J=5.5 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.5, 47.5, 48.4, 51.1, 63.7, 72.3, 96.9,126.9 (2×C), 131.0, 133.6 (2×C), 139.5 (2×C), 143.2 (2×C), 161.1, 163.8.

MS-ESI m/z (% rel. Int.): 451.9 ([MH]⁺, 100), 363.8 (45), 342.9 (70),216.9 (75), 148.0 (30).

HPLC: Method A, detection UV 254 nm, Compound 225 RT=3.83 min, peak area98.7%.

Preparation of(±)-threo-2-(2-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 226

To a stirred solution of(±)-threo-2-amino-3-hydroxy-3-pyridin-4-yl-1-pyrrolidin-1-yl-propan-1-oneCompound 22 (500 mg, 1.62 mmol) in 10 ml methanol was added successivelyEt₃N (496 μL, 3.57 mmol) and 2-iodobenzaldehyde (414 mg, 1.78 mmol) in 1ml methanol. The mixture was stirred 6 h at RT under nitrogen and aceticacid (464 μL, 8.11 mmol) and sodium cyanoborohydride (356 mg, 5.57 mmol)were added. The mixture was stirred overnight at 20° C. and evaporatedto give a residue which was partitioned between CH₂Cl₂ and 1 N aqueouspotassium hydroxyde. The organic layer was washed with brine and driedover MgSO₄, filtered and evaporated. The crude product was purified bycolumn chromatography (SiO₂, EtOAc:MeOH=9:1). After evaporation anddrying a white solid(±)-threo-2-(2-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onewas obtained (419.6 mg, 57% yield). The product was dissolved inmethanol (10 mL). The solution was stirred at room temperature and asolution of HCl (1 M, 7.4 mL) was added via syringe at RT for 10 min.The mixture was concentrated and triturated with EtOAc. After filtrationand drying,(±)-threo-2-(2-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 226 was obtained (195 mg, 25% yield) as a whitesolid.

MW: 524.22; Yield: 25%; White Solid; Mp (° C.): 148.0

R_(f): 0.50 (MeOH:EtOAc=20:80, free base).

¹H-NMR (CD₃OD, δ): 1.33-1.80 (m, 4H, 2×CH₂), 2.12-2.25 (m, 1H, CH₂),2.75-2.88 (m, 1H, CH₂), 3.14-3.25 (m, 1H, CH₂), 3.25-3.31 (m, 1H, CH₂),4.14 (d, 1H, J=8.4 Hz, NH—CH), 4.37-4.52 (m, 2H, NH—CH₂) 5.10 (d, 1H,J=8.3 Hz, CH—O), 7.18 (t, 1H, J=7.6 Hz, ArH), 7.49 (d, 1H, J=7.5 Hz,ArH), 7.61 (d, 1H, J=7.7 Hz, ArH), 7.68 (d, 2H, J=5.2 Hz, ArH), 7.97 (d,J=7.9 Hz, 1H, ArH), 8.66 (d, J=4.9 Hz, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.5, 47.6, 47.8, 55.3, 64.3, 72.7, 101.8,124.4 (2×C), 130.3, 132.0, 132.5, 135.1, 141.7 (2×C), 148.2, 154.0,164.2.

MS-ESI m/z (% rel. Int.): 451.9 ([MH]⁺, 100), 352.9 (55), 342.9 (30).

HPLC: Method A, detection UV 254 nm, Compound 226 RT=3.72 min, peak area98.95%.

Preparation of(±)-threo-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 227

To a stirred solution of(±)-threo-2-amino-3-hydroxy-3-pyridin-4-yl-1-pyrrolidin-1-yl-propan-1-oneCompound 22 (500 mg, 1.62 mmol) in 10 ml methanol was added successivelyEt₃N (496 μL, 3.55 mmol) and 3-iodobenzaldehyde (414 mg, 1.78 mmol) in 1ml methanol. The mixture was stirred 6 h at RT under nitrogen and aceticacid (464 μL, 8.11 mmol) and sodium cyanoborohydride (356 mg, 5.57 mmol)were added. The mixture was stirred overnight at 20° C. then evaporatedand the residue was partitioned between CH₂Cl₂ and 1 N aqueous potassiumhydroxyde. The organic layer was washed with brine and dried. Theorganic layer was evaporated. The crude product was purified by columnchromatography (SiO₂, CH₂Cl₂:MeOH=95:5). After evaporation and drying awhite solid(±)-threo-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-one(437 mg, 60% yield). This product was dissolved in methanol (10 mL). Thesolution was stirred at RT and a solution of HCl (1 M, 7.7 mL) was addedat room temperature for 10 min. The mixture was concentrated andtriturated with diethyl ether. After filtration and drying,(±)-threo-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 227 was obtained (205 mg, 26% yield) as a whitesolid.

MW: 524.22; Yield: 26%; White Solid; Mp (° C.): 142.8

R_(f): 0.30 (MeOH:CH₂Cl₂=5:95, free base).

¹H-NMR (CD₃OD, δ): 1.25-1.80 (m, 4H, 2×CH₂), 2.03-2.12 (m, 1H, CH₂),2.80-2.92 (m, 1H, CH₂), 3.11-3.25 (m, 2H, CH₂), 4.06-4.30 (m, 3H, CH—N &N—CH₂), 5.04 (d, 1H, J=8.8 Hz, CH—OH), 7.23 (t, 1H, J=7.8 Hz, ArH), 7.51(d, 1H, J=7.5 Hz, ArH), 7.67 (d, 2H, J=5.8 Hz, ArH), 7.80 (d, 1H, J=5.2Hz, ArH), 7.87 (s, 1H, ArH), 8.66 (d, J=5.2 Hz, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.5, 47.3, 47.7, 50.5, 64.3, 72.8, 95.3,124.5 (2×C), 130.9, 132.0, 133.9, 140.1, 140.5, 148.3, 153.7, 164.2.

MS-ESI m/z (% rel. Int.): 451.9 ([MH]⁺, 100), 352.9 (40), 342.9 (50).

HPLC: Method A, detection UV 254 nm, Compound 227 RT=3.80 min, peak area98.7%.

Preparation of(+)-(2S,3R)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 228

To a stirred solution of(+)-(2S,3R)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 204 (204 mg, 0.67 mmol) in MeOH (6 mL) wasadded Et₃N (202 μL, 1.46 mmol) and, via syringe, a solution of3-iodobenzaldehyde (169 mg, 0.73 mmol) in methanol (1 mL). The mixturewas stirred 5 h at RT under nitrogen. CH₃COOH (190 μL, 3.30 mmol) andsodium cyanoborohydride (146 mg, 2.32 mmol) were added and the reactionmixture was stirred overnight at RT. MeOH was evaporated and the residuewas partitioned between CH₂Cl₂ and a solution of 1 N aqueous potassiumcarbonate. The organic layer was washed with brine, dried over MgSO₄,filtered and evaporated. The crude product was purified by columnchromatography (SiO₂, CH₂Cl₂:MeOH=95:5). After evaporation and drying(2S,3R)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-one(246 mg) was obtained as a white solid. The product was dissolved inmethanol (5 mL) and a solution of HCl in MeOH (1 M, 2.5 mL) was addedvia syringe and the solution was stirred at RT for 0.6 h. The mixturewas concentrated and the resulting solid triturated with Et₂O. Afterfiltration and drying(+)-(2S,3R)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 228 (280 mg, 80% yield) was obtained as a whitesolid.

MW: 524.22; Yield: 80%; White Solid; Mp (° C.): 107.4

α²² _(D)=+101.9 (MeOH, c=1.02).

R_(f): 0.30 (MeOH:CH₂Cl₂=5:95, free base).

¹H-NMR (CD₃OD, *): 1.52-1.82 (m, 4H, 2×CH₂), 2.38-2.45 (m, 1H, CH₂),3.01-3.09 (m, 1H, CH₂), 3.18-3.32 (m, 2H, CH₂), 4.20-4.29 (dd, 2H,J=28.1 Hz, J=13.4 Hz, NH—CH₂), 4.40 (d, 1H, J=7.9 Hz, N—CH), 5.30 (d,J=7.9 Hz, 1H, CH—O), 7.25 (t, 1H, J=5.4 Hz, ArH), 7.54 (d, 1H, J=7.1 Hz,ArH), 7.81 (d, 1H, J=7.2 Hz, ArH), 7.89 (d, 1H, J=1.3 Hz, ArH), 8.13 (d,J=5.5 Hz, 2H, ArH), 8.90 (d, J=5.3 Hz, 2H, ArH).

MS-ESI m/z (% rel. Int.): 452.1 ([MH]⁺, 100), 353.0 (65), 343.1 (80).

HPLC: Method A, detection UV 254 nm, Compound 228 RT=3.87 min, peak area97.0%.

Preparation of(2R,3S)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 229

Similar to Compound 228 with(−)-(2R,3S)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 203 (77 mg, 0.25 mmol) in 2.3 ml of MeOH, Et₃N(76 μL, 0.79 mmol), 3-iodobenzaldehyde (64 mg, 0.275 mmol), CH₃COOH(78.6 μL, 1.37 mmol) and NaBH₃CN (60.5 mg, 0.96 mmol).

(2R,3S)-2-(3-iodobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)-propan-1-onedihydrochloride Compound 229 (70 mg, 53.5% yield) was obtained as a paleyellow solid.

MW: 524.22; Yield: 53.5%; Pale Yellow Solid; Mp (° C.): 179.7

R_(f): 0.30 (MeOH:CH₂Cl₂=5:95, free base).

¹H-NMR (CD₃OD, *): idem to Compound 228.

MS-ESI m/z (% rel. Int.): 452.1 ([MH]⁺, 100), 353.0 (70), 343.1 (60).

HPLC: Method A, detection UV 254 nm, Compound 228 RT=3.78 min, peak area99.0%.

Preparation of(±)-threo-2-amino-3-hydroxy-1-morpholino-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 230trans-(4,5-Dihydro-5-(thiophen-3-yl)oxazol-4-yl)(morpholino)methanoneSLA 09052A

SLA 09052A was prepared in accordance with method D usingthiophene-3-carbaldehyde (0.768 mL, 5.35 mmol), KOH (0.273 mg, 4.86mmol) in methanol (5 mL) and 2-isocyano-1-morpholinoethanone SLA 07118(0.75 g, 4.86 mmol). The solution was stirred for 2 h at 0° C. Afterwork-up the residue was purified by column chromatography (florisil,EtOAc). After evaporation and drying,trans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)(morpholino)methanoneSLA 09052A (0.327 g, 25% yield) was obtained as a yellow oil.

MW: 266.32; Yield: 25%; Yellow Oil.

¹H-NMR (CDCl₃, *): 3.40-4.00 (m, 8H, 4×CH₂), 4.67 (dd, 1H, J=7.3 Hz,J=2.2 Hz, CH—N), 6.29 (d, 1H, J=7.3 Hz, CH—O), 6.97 (d, 1H, J=2.2 Hz,CH═N), 7.01 (dd, 1H, J=5.0 Hz, J=1.3 Hz, CH═C), 7.28-7.40 (m, 2H, CH═C).

(±)-threo-2-Amino-3-hydroxy-1-morpholino-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 230

To a solution oftrans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)(morpholino)methanoneSLA 09052A (0.327 g, 1.12 mmol) in MeOH (5 mL) was added HCl 37% (2 mL).After heating (50° C.) the mixture for 24 h the reaction mixture wasconcentrated and the crude product was coevaporated twice with EtOAc.After trituration with EtOAc, filtration and drying(±)-threo-2-amino-3-hydroxy-1-morpholino-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 230 (276 mg, 77% yield) was obtained as a paleyellow solid.

MW: 292.78; Yield: 77%; Pale Yellow Solid; Mp (° C.): 209.1

¹H-NMR (CD₃OD, δ): 2.77-2.92 (m, 2H, CH₂), 3.20-3.60 (m, 6H, 3×CH₂),4.49 (d, 1H, J=8.7 Hz, CH—N), 4.95 (d, 1H, J=8.8 Hz, CH—O), 7.18 (d, 1H,J=4.5 Hz, CH═C), 7.44 (d, 1H, J=1.7 Hz CH═C), 7.44 (dd, 1H, J=4.5 Hz,J=1.7 Hz CH═C).

Preparation of(±)-threo-2-amino-3-hydroxy-1-(piperidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 231trans-(4,5-Dihydro-5-(thiophen-3-yl)oxazol-4-yl)(piperidin-1-yl)methanoneSLA 09052B

SLA 09052B was prepared in accordance with method D usingthiophene-3-carbaldehyde (0.778 mL, 5.43 mmol), KOH (0.273 mg, 4.94mmol) in methanol (5 mL) and 2-isocyano-1-(piperidin-1-yl)ethanone SLA07116B (0.75 g, 4.94 mmol). The solution was stirred for 2 h at 0° C.After work-up (without any further purification) and dryingtrans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)(piperidin-1-yl)methanoneSLA 09052B was obtained as a yellow oil (1.29 g, 99% yield).

MW: 264.34; Yield: 99%; Yellow Oil.

¹H-NMR (CDCl₃, δ): 1.47-1.75 (m, 6H, 3×CH₂), 3.42-3.82 (m, 4H, 2×CH₂),4.72 (dd, 1H, J=7.2 Hz, J=2.2 Hz, CH—N), 6.28 (d, 1H, J=7.2 Hz, CH—O),6.96 (d, 1H, J=2.2 Hz, CH═N), 7.01 (dd, 1H, J=5.0 Hz, J=1.2 Hz, CH═C),7.30-7.35 (m, 2H, CH═C).

¹³C-NMR (CDCl₃, 6): 24.5, 25.5, 26.5, 43.7, 46.8, 73.4, 78.0, 122.6,125.1, 127.2, 140.5, 155.0, 166.3.

(±)-threo-2-Amino-3-hydroxy-1-(piperidin-1-yl)-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 231

To a solution oftrans-(4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)(piperidin-1-yl)methanoneSLA 09052B (1.29 g, 4.88 mmol) in methanol (5 mL) was added hydrochloricacid 37% (5 mL). After heating (50° C.) the mixture for 24 h thereaction mixture was concentrated and the crude product was coevaporatedtwice with EtOAc. After trituration with EtOAc, filtration and drying(±)-threo-2-amino-3-hydroxy-1-morpholino-3-(thiophen-3-yl)propan-1-onehydrochloride Compound 231 was obtained as a pale yellow solid (1.07 g,75% yield).

MW: 290.81; Yield: 75%; Pale Yellow Solid; Mp (° C.): 210.6

¹H-NMR (CD₃OD, δ): 0.82-0.97 (m, 1H, 0.5×CH₂), 1.30-1.70 (m, 5H,2.5×CH₂), 2.75-2.91 (m, 1H, CH₂), 3.12-3.55 (m, 3H, 1.5×CH₂), 4.51 (d,1H, J=8.4 Hz, CH—N), 4.94 (d, 1H, J=8.5 Hz, CH—O), 7.16 (d, 1H, J=4.9Hz, CH═C), 7.44 (d, 1H, J=2.5, Hz CH═C), 7.44 (dd, 1H, J=4.5 Hz, J=3.2Hz CH═C).

Preparation of(±)-threo-2-amino-3-hydroxy-1-morpholino-3-(pyridin-3-yl)propan-1-onedihydrochloride Compound 232trans-(4,5-Dihydro-5-(pyridin-3-yl)oxazol-4-yl)(morpholino)methanone SLA09050A

SLA 09050A was prepared in accordance with method D usingpyridine-3-carbaldehyde (0.65 mL, 4.86 mmol), KOH (0.273 mg, 4.86 mmol)in methanol (10 mL) and 2-isocyano-1-morpholinoethanone SLA 07118 (0.75g, 4.86 mmol). The solution was stirred for 20 h at 0° C. After work-up(without any further purification), evaporation and dryingtrans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(morpholino)methanone SLA09050A was obtained as a yellow solid (0.92 g, 72.5% yield).

MW: 261.28; Yield: 72.5%; Yellow Solid.

¹H-NMR (CDCl₃, δ): 3.42-4.00 (m, 8H, 4×CH₂), 4.63 (dd, 1H, J=7.7 Hz,J=2.3 Hz, CH—N), 6.29 (d, 1H, J=7.7 Hz, CH—O), 7.02 (d, 1H, J=2.2 Hz,CH═N), 7.33 (m, 1H, ArH), 7.60-7.66 (m, 1H, ArH), 8.57-8.62 (m, 2H,ArH).

¹³C-NMR (CDCl₃, 6): 43.0, 46.3, 66.7, 66.8, 74.6, 79.2, 123.5, 133.4,135.1, 147.5, 150.0, 155.0, 166.3.

MS-ESI m/z (% rel. Int.): 262.1 ([MH]⁺, 55), 108.0 (100).

(±)-threo-2-Amino-3-hydroxy-1-morpholino-3-(pyridin-3-yl)propan-1-onedihydrochloride Compound 232

To a solution oftrans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(morpholino)methanone SLA09050A (0.911 g, 3.48 mmol) in methanol (10 mL) was added hydrochloricacid 37% (5 mL). After heating (50° C.) the mixture for 2.25 h thereaction mixture was concentrated and the crude product was coevaporatedtwice with EtOAc and the crude product was dissolved in a 1 N solutionof K₂CO₃ which was extracted with a mixture CH₂Cl₂:iPrOH=9:1 (6×100 mL).The combined organic layer was dried over MgSO₄, filtered and evaporatedto led to a crude product which was purified by column chromatography(SiO₂, EtOAC:MeOH=70:30). After evaporation the product was dissolved ina solution of HCl in MeOH (0.5M, 29 mL) and stirred at RT for 1.5 h. Theproduct was co-evaporated twice with EtOAc. After trituration withEtOAc, filtration and drying,(±)-threo-2-amino-3-hydroxy-1-morpholino-3-(pyridin-3-yl)propan-1-onedihydrochloride Compound 232 (270 mg, 24% yield) was obtained as ayellow solid.

MW: 324.2; Yield: 24%; Yellow Solid.

¹H-NMR (CD₃OD, δ): 3.20-3.78 (m, 8H, 4×CH₂), 4.87 (d, 1H, J=5.5 Hz,CH—N), 5.40 (d, 1H, J=5.4 Hz, CH—O), 8.20 (m, 1H, ArH), 8.77 (d, 1H,J=8.3 Hz, ArH), 8.93 (d, 1H, J=5.5 Hz, ArH), 9.03 (s, 1H, ArH).

Preparation of(±)-threo-2-amino-3-hydroxy-1-piperidin-3-(pyridin-3-yl)propan-1-onedihydrochloride Compound 233trans-(4,5-Dihydro-5-(pyridin-3-yl)oxazol-4-yl)(piperidin)methanone SLA09050B

SLA 09050B was prepared in accordance with method D usingpyridine-3-carbaldehyde (0.512 mL, 5.42 mmol), KOH (0.277 mg, 4.93 mmol)in methanol (10 mL) and 2-isocyano-1-piperidinethanone SLA 07116B (0.75g, 4.93 mmol). The solution was stirred for 20 h at 0° C. After work-up(without any further purification), evaporation and drying,trans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(piperidin)methanone SLA09050B (0.917 g, 72% yield) was obtained as a yellow solid.

MW: 259.30; Yield: 72%; Yellow Solid.

¹H-NMR (CDCl₃, δ): 1.59-1.73 (m, 6H, 3×CH₂), 3.44-3.83 (m, 4H, 2×CH₂),4.68 (dd, 1H, J=7.6 Hz, J=2.3 Hz, CH—N), 6.30 (d, 1H, J=7.6 Hz, CH—O),7.02 (d, 1H, J=2.2 Hz, CH═N), 7.32 (m, 1H, ArH), 7.60-7.66 (m, 1H, ArH),8.57-8.62 (m, 2H, ArH).

¹³C-NMR (CDCl₃, δ): 24.4, 25.5, 26.4, 43.8, 46.9, 74.6, 79.5, 123.6,133.5, 135.4, 147.5, 149.9, 154.8, 165.9.

(±)-threo-2-Amino-3-hydroxy-1-piperidin-3-(pyridin-3-yl)propan-1-onedihydrochloride Compound 233

To a solution oftrans-(4,5-dihydro-5-(pyridin-3-yl)oxazol-4-yl)(piperidin)methanone SLA09050B (0.917 g, 3.54 mmol) in methanol (10 mL) was added hydrochloricacid 37% (5 mL). After heating at 50° C. the mixture for 2.25 h thereaction mixture was concentrated and the crude product was coevaporatedtwice with EtOAc. After trituration with EtOAc, filtration and drying,the crude product was dissolved in a 1 N solution of K₂CO₃ and theproduct was extracted CH₂Cl₂:iPrOH=9:1 (6×100 mL). The crude product waspurified by column chromatography (EtOAc:MeOH=7:3)(±)-threo-2-amino-3-hydroxy-1-piperidin-3-(pyridin-3-yl)propan-1-one wasobtained pale yellow solid (223.5 mg). The product was dissolved in asolution of HCl in MeOH (0.5 M, 18 mL) and stirred at RT for 1.5 h.After trituration with EtOAc, filtration and drying,(±)-threo-2-amino-3-hydroxy-1-piperidin-3-(pyridin-3-yl)propan-1-onedihydrochloride Compound 233 (208 mg, 18% yield) was obtained as ayellow solid.

MW: 322.2; Yield: 18%; Yellow Solid.

¹H-NMR (CD₃OD, *): 1.15-1.80 (m, 6H, 4×CH₂), 3.10-3.80 (m, 4H, 2×CH₂),4.88 (d, 1H, J=5.6 Hz, CH—N), 5.33 (d, 1H, J=5.6 Hz, CH—O), 8.19 (t, 1H,J=7.1 Hz, ArH), 8.74 (d, 1H, J=7.9 Hz, ArH), 8.93 (d, 1H, J=5.6 Hz,ArH), 9.01 (s, 1H, ArH).

Preparation of(2R,3S)-2-((R)-2-hydroxy-1-phenylethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 234 (R)-5-Phenylmorpholin-2-one EBE 06134

To a solution of phenylbromoacetate (18.58 g, 86 mmol) in CH₃CN undernitrogen was added a solution of (R)-phenylglycinol (10.17 g, 74 mmol)and di-isopropylethylamine (34 mL, 195 mmol) in CH₃CN. The volatileswere removed under reduced pressure keeping the bath temperature below25° C. to obtain an oil that was treated with EtOAc (120 mL) and stirredfor 15 min. The resulting white precipitate was removed by filtration.The filtrate was concentrated under reduced pressure and the desiredproduct was isolated using column chromatography (SiO₂) with a stepgradient of 10% to 50% [v/v] EtOAc in cyclohexane to give afterevaporation (R)-5-phenylmorpholin-2-one EBE 06134 (3.17 g, 24% yield) asa white solid.

MW: 177.2; Yield: 24%; White Solid; Mp (° C.): 50.3

R_(f): 0.30 (EtOAc:cyclohexane=50:50).

¹H-NMR (CDCl₃, δ): 1.99 (s, 1H, NH), 3.89 (q, 2H, J=17.8 Hz, N—CH₂),4.18 (dd, 1H, J=3.7 Hz, J=10.3 Hz, O—CH), 4.29 (t, 1H, J=10.5 Hz, N—CH),4.40 (dd, 1H, J=3.7 Hz, J=10.5 Hz, O—CH), 7.30-7.45 (m, 5H, ArH).

¹³C-NMR (CDCl₃, δ): 46.8, 54.8, 72.8, 125.3 (2×C), 127.0, 127.3 (2×C),135.9, 166.0.

[α]²² _(D)=+30.3° (c=1.00, MeOH).

(1S,3R,5R,8aR)-Tetrahydro-5-phenyl-1,3-di(pyridin-4-yl)oxazolo[4,3-c][1,4]oxazin-8(3H)-oneEBE 06136

A solution of (R)-5-phenylmorpholinon-2-one EBE 06134 (3.0 g, 16.9 mmol)and pyridine-4-carboxaldehyde (5.43 g, 50.7 mmol) in toluene (75 mL) wasrefluxed in a soxhlet extractor filled with molecular sieves 4A (25 g)for 16 hours. All the volatiles were evaporated and the desired productwas purified by column chromatography (SiO₂) using a gradient of 80% to100% [v/v] EtOAc in cyclohexane to give after evaporation(1S,3R,5R,8aR)-tetrahydro-5-phenyl-1,3-di(pyridin-4-yl)oxazolo[4,3-c][1,4]oxazin-8(3H)-oneEBE 06136 (1.7 g, 46% yield) as a pale yellow solid.

MW: 373.4; Yield: 46%; Pale Yellow Solid; Mp (° C.): 155.6

R_(f): 0.20 (EtOAc).

¹H-NMR (CDCl₃, δ): 4.10-4.17 (m, 1H, N—CH), 4.25 (dd, 1H, J=3.3 Hz,N—CH), 4.36-4.54 (m, 2H, O—CH), 5.38 (d, 1H, O—CH, J=8.2 Hz), 5.53 (s,1H, N—CH), 7.20-7.35 (m, 8H, ArH), 7.40-7.50 (m, 1H, ArH), 8.51 (dd, 2H,J=1.4 Hz, J=4.5 Hz), 8.57 (dd, 2H, J=1.4 Hz, J=4.5 Hz, ArH).

((2R,4S,5R)-3-((R)-2-Hydroxy-1-phenylethyl)-2,5-di(pyridin-4-yl)oxazolidin-4-yl)(pyrrolidin-1-yl)methanoneEBE 06138

To a solution of(1S,3R,5R,8aR)-tetrahydro-5-phenyl-1,3-di(pyridin-4-yl)oxazolo[4,3-c][1,4]oxazin-8(3H)-oneEBE 06136(1.7 g, 4.55 mmol) in CH₂Cl₂ was added pyrrolidine (1.90 mL,22.8 mmol) and the solution was stirred under nitrogen at 25° C. for 16h. All the volatiles were evaporated and the resulting product wasisolated using column chromatography (SiO₂) with a gradient of 0-20%[v/v] MeOH in EtOAc to give after evaporation((2R,4S,5R)-3-((R)-2-hydroxy-1-phenylethyl)-2,5-di(pyridin-4-yl)oxazolidin-4-yl)(pyrrolidin-1-yl)methanoneEBE 06138 (0.665 g, 33% yield) as a white solid.

MW: 444.5; Yield: 33%; White Solid; Mp (° C.): 63.6.

R_(f): 0.25 (MeOH:EtOAc=20:80).

¹H-NMR (CDCl₃, δ); 1.70-1.90 (m, 4H, 2×CH₂), 2.25 (bs, 1H, OH),2.75-2.85 (m, 1H, CH—N), 2.95-3.05 (m, 1H, N—CH), 3.50-3.60 (m, 2H,N—CH₂), 3.80-4.15 (m, 4H, 2×CH+CH₂—O), 5.10 (d, 1H, J=4.7 Hz, CH), 6.32(s, 1H, CH), 7.18-7.32 (m, 7H, ArH), 7.45 (d, 2H, J=5.9 Hz, ArH), 8.54(d, 2H, J=6.0 Hz, ArH), 8.64 (d, 2H, J=6.0 Hz).

¹³C-NMR (CDCl₃, 6): 23.9, 26.0, 46.3, 46.6, 60.3, 63.8, 65.2, 94.1,121.1 (2×C), 123.0 (2×C), 128.0, 128.3 (2×C), 128.5 (2×C), 137.8, 147.8.MS-ESI m/z (% rel. Int.): 445.1 ([MH]⁺, 20).

HPLC: Method A, detection at 254 nm, EBE 06138 RT=3.50 min, peak area99%.

[α]²² _(D)=−16.0° (c=1.00, CHCl₃).

(2R,3S)-2-((R)-2-Hydroxy-1-phenylethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 234

To a solution of((2R,4S,5R)-3-((R)-2-hydroxy-1-phenylethyl)-2,5-di(pyridin-4-yl)oxazolidin-4-yl)(pyrrolidin-1-yl)methanoneEBE 06138 (600 mg, 1.34 mmol) in MeOH (6 mL) was added a solution of 1 NHCl (6 mL) and the reaction mixture was stirred at 60° C. for 2 h,evaporated to dryness to give a white solid (745 mg). This crude productwas dissolved in CH₂Cl₂ (10 mL) and Na₂CO₃ (sat. sol.). The organiclayer was separated was separated and the aqueous layer was washed withCH₂Cl₂ (5×2 mL). The combined organic layer were dried over Na₂SO₄,filtered over cotton mixed with silica gel (600 mg), evaporated andloaded on a silica gel column of 25 g. The desired product was elutedusing a gradient of MeOH 0% to 20% in EtOAc to give after evaporation(2R,3S)-2-((R)-2-hydroxy-1-phenylethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-one(320 mg, 67% yield) as a white solid.

MW=355.43; Yield: 67%; White Solid; Mp (° C.)=76.4

R_(f): 0.3 (MeOH:EtOAc=20:80).

¹H NMR (CDCl₃, δ): 1.18-1.30 (m, 2H, CH₂), 1.30-1.48 (m, 2H, CH₂),1.80-1.90 (m, 1H, CH₂), 2.23-2.33 (m, 1H, N—CH₂), 2.85-2.95 (m, 2H,N—CH₂), 3.00-3.12 (m, 2H, N—CH+N—CH), 3.72-3.85 (m, 3H, O—CH₂+NH), 4.58(d, 1H, J=8.4 Hz, O—CH), 7.18-7.35 (m, 7H, ArH), 8.51 (d, 2H, J=6.0 Hz,ArH).

[α]²² _(D)=−68.8° (c=1.00, CHCl₃).

To a solution of(2R,3S)-2-((R)-2-hydroxy-1-phenylethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onein MeOH (100 mg), in MeOH (1 mL) was added a solution HCl (1 N, 1.1 mL)at 0° C. and the solution was stirred for 10 min to give afterevaporation(2R,3S)-2-((R)-2-hydroxy-1-phenylethylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 234 (120 mg, 99% yield) as a white solid.

MW: 430.4; Global Yield: 66%; White Solid; Mp (° C.): 38.6.

¹H-NMR (CD₃OD, δ): 1.38-1.58 (m, 4H, CH₂), 2.12-2.22 (m, 1H, N—CH₂),2.80-2.90 (m, 1H, N—CH₂), 3.00-3.10 (m, 2H, N—CH₂), 3.91 (dd, 1H, J=4.3Hz, J=11.3 Hz, CH₂—O), 4.02-4.12 (m, 1H, CH₂—O), 4.55-4.65 (m, 2H,N—CH), 5.27 (d, 1H, J=8.9 Hz, 7.35-7.45 (m, 3H, ArH), 7.45-7.58 (m, 2H,ArH).

¹³C-NMR (CD₃OD, δ): 24.5, 26.3, 47.2, 48.0, 63.5, 64.4, 67.3, 72.6,126.7, 130.2, 131.5, 132.3, 143.3, 160.7, 163.8.

Preparation ofN-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamidehydrochloride Compound 235

To a suspension of(±)-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (300 mg, 0.89 mmol) in CH₂Cl₂ (6 mL) wasadded Et₃N (370 μL, 2.67 mmol) and the mixture was stirred for 10 min,cooled at 4° C. and a solution of acetic anhydride (65 mL, 0.89 mmol) inCH₂Cl₂ was added dropwise for 10 min. The reaction mixture was broughtto room temperature, stirred for 16 h and washed with water (3×4 mL),NaOH (0.5 N) (3×4 mL) evaporated to give an oily residue that waspurified using column chromatography (SiO₂) with a gradient of 0% to 20%MeOH in EtOAc. The product in MeOH at 4° C. was treated with a solutionof 1 N HCl (4 mL) and all the volatiles were evaporated. Product wasprecipitated using a mixture of methanol in EtOAc to obtain afterevaporationN-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)acetamidehydrochloride Compound 235 (209 mg, 52% yield).

MW: 313.78; Yield: 52%; White Solid; Mp (° C.): 181.3

R_(f): 0.20 (MeOH:EtOAc=20:80, free base).

¹H-NMR (CD₃OD, δ): 1.82-2.05 (m, 7H, 2×CH₂ & CH₃), 3.35-3.45 (m, 2H,CH₂), 3.50-3.65 (m, 2H, CH₂), 5.10 (d, 1H, J=3.8 Hz, N—CH), 5.39 (d, 1H,J=3.9 Hz, O—CH), 8.16 (d, 2H, J=6.2 Hz, ArH), 8.81 (d, 2H, J=6.7 Hz,ArH).

¹³C-NMR (CD₃OD, δ): 22.1, 25.0, 27.0, 47.5, 48.2, 57.0, 72.0, 126.6(2×C), 142.0 (2C), 142.0 (2×C), 164.4, 169.0, 173.0.

Preparation of(R)-2-amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 236 tert-Butyl(R)-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamate EBE06172

N-Boc-(S)-3-(pyridin-4-yl)alanine (0.500 g, 1.88 mmol) was dissolved inCH₂Cl₂ (15 mL) and DIEA (361 μL, 2.07 mmol) was added. The mixture wascooled to 0° C. and isobutyl chloroformate (270 μL, 2.07 mmol) wasadded. The mixture was stirred for 10 min and pyrrolidine (267 mg, 3.76mmol) in CH₂Cl₂ (5 mL) was added. This mixture was stirred for 15 min at4° C., 12 h at 25° C., washed successively with NaH₂PO₄, saturatedsodium hydrogen carbonate, water and brine. The organic layer was driedover magnesium sulfate and evaporated to dryness. The residue obtainedwas purify by column chromatography (SiO₂) using a gradient of MeOH0-10% [v/v] in EtOAc to give tert-butyl(R)-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamate EBE06172 (146 mg, 24% yield) as a white solid.

MW: 319.4; Yield: 24%; White Solid; Mp (° C.): 52.5.

R_(f): 0.30 (MeOH:EtOAc=20:80).

¹H-NMR (CDCl₃, δ): 1.40 (s, 9H, (CH₃)₃), 1.65-1.90 (m, 4H, CH₂),2.80-2.90 (m, 1H, CH), 2.90-3.00 (m, 2H, CH₂), 3.28-3.38 (m, 1H, N—CH),3.40-3.50 (m, 2H, N—CH₂), 4.60-4.70 (m, 1H, N—CH), 5.38-5.48 (m, 1H,NH), 7.16 (d, 2H, J=4.5 Hz, ArH), 8.50 (d, 2H, J=4.5 Hz, ArH).

MS-ESI m/z (% rel. Int.): 320.2 ([MH]⁺, 20).

HPLC: Method A, detection at 254 nm, EBE 06172 RT=3.82 min, peak area85%.

(R)-2-Amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 236

To a solution of TFA (2 mL) in CH₂Cl₂ (8 mL) was added(R)-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamate EBE06172 (146 mg, 0.654 mmol) and the mixture was stirred for 2 h at 25° C.The volatiles were evaporated and the product was treated with asuspension on amberlite-400 (OH⁻ form, 2 g) in MeOH. The suspension wasfiltered and washed with MeOH (3×5 mL). The combined methanol fractionswere evaporated under reduced pressure and the desired product wasisolated using column chromatography (SiO₂) with a mixture ofEtOAc:MeOH:NH₄OH=70:30:4 to give an oily residue that was treated with asolution of 0.1 N HCl in iPrOH for 10 min. Evaporation of the volatileafforded (R)-2-amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 236 (78 mg, 47% yield) as a pale yellow solid.

MW: 255.75; Yield: 47%; Pale Yellow Solid; Mp (° C.): 127.5

R_(f): 0.30 (EtOAc:MeOH:NH₄OH=70:30:4, free base).

¹H-NMR (CD₃OD, δ): 1.80-2.05 (m, 4H, CH₂), 3.28-3.80 (m, 6H, CH₂,N—CH₂), 3.70 (t, 1H, J=6.7 Hz, CH—N), 8.10 (d, 2H, J=5.9 Hz, ArH), 8.88(d, 2H, J=5.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 23.4, 25.4, 35.6, 46.1, 46.5, 51.1, 128.5 (2×C),146.3 (2×C), 156.1, 165.0.

Preparation of tert-butyl5-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamoyl)pentylcarbamateCompound 237

To a solution of N-Boc-aminohexanoic acid (342 mg, 1.48 mmol) in THF (10mL) was added N-methylmorpholine (163 μL, 1.48 mmol). The solution wasstirred for 5 min, cooled at −15° C. and treated dropwise with isobutylchloroformate (211 μL, 1.48 mmol). This solution was added via astainless steal cannula to a solution of(±)-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 22 (500 mg, 1.48 mmol) and N-methyl-morpholine(489 mg, 1.47 mmol) in THF (10 mL) at −15° C. The reaction mixture waskept for 0.5 h at −15° C. followed by 2 h at 25° C. with continuousstirring. After evaporation of the solvent, the residue was partitionedbetween EtOAc and H₂O, washed with NaH₂PO₄, saturated aqueous NaHCO₃,dried over sodium sulfate and purified by column chromatography (SiO₂)with a gradient of 0% to 10% [v/v] MeOH in EtOAc to give tert-butyl5-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamoyl)pentylcarbamateCompound 237 (455 mg, 69% yield) as a white solid.

MW: 448.6; Yield: 69%; White Solid.

R_(f): 0.20 (EtOAc:MeOH=90:10).

¹H-NMR (CD₃OD, δ): 1.05-1.15 (m, 2H, CH₂), 1.35-1.55 (m, 13H,2×CH₂+C(CH₃)₃), 1.75-1.95 (m, 4H, 2×CH₂), 2.00-2.20 (m, 2H, O═CCH₂),3.05 (q, 2H, J=6.7 Hz, N—CH₂), 3.20-3.35 (m, 1H, N—CH), 3.38-3.50 (m,2H, N—CH₂), 3.65-3.75 (m, 1H, N—CH), 4.72 (bs, 1H, NH), 4.98 (dd, 1H,J=8.8 Hz, J=3.6 Hz), 5.08 (d, 1H, J=3.3 Hz, OCH), 5.23 (bs, 1H, OH),6.50 (d, 1H, J=8.7 Hz, NH), 7.35 (d, 2H, J=6.0 Hz, ArH), 8.58 (d, 2H,J=4.6 Hz, J=1.4 Hz, ArH).

MS-ESI m/z (% rel. Int.): 449.2 ([MH]⁺, 30), 349.2 (100).

HPLC: Method A, detection at 254 nm, Compound 237 RT=4.03 min, peak area99.9%.

Preparation of6-amino-N-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)hexanamideCompound 238

To a solution of tert-butyl5-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-ylcarbamoyl)pentylcarbamateCompound 237 (81 mg, 0.181 mmol) in CH₂Cl₂ (8 mL) was added TFA (2 mL)at 0° C. and stirred for 2 h at 0° C. All the volatiles were evaporatedto give a residue that was treated with a suspension of Amberlite-400(OH⁻) in MeOH. After filtration, the filtrate was evaporated and theproduct was isolated by column chromatography (SiO₂) withCH₂Cl₂:MeOH:NH₄OH=10:5:0.4 to afford6-amino-N-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)hexanamideCompound 238 (40 mg, 64% yield) as a white solid.

MW: 448.6; Yield: 64%; White Solid; Mp (° C.): 134.4

R_(f): 0.30 (CH₂Cl₂:MeOH:NH₄OH=10:5:0.4).

¹H NMR (CDCl₃, δ): 1.12-1.30 (m, 2H, CH₂), 1.30-1.50 (m, 2H, CH₂),1.50-1.65 (m, 2H, CH₂), 1.65-1.95 (m, 4H, CH₂), 2.10-2.30 (m, 2H, CH₂),2.55-2.70 (t, 2H, J=6.9 Hz, CH₂), 3.10-3.20 (m, 2H, CH₂), 3.28-3.50 (m,2H, CH₂), 3.60-3.70 (m, 1H, CH), 4.95 (dd, 1H, J=5.1 Hz, J=8.4 Hz,O—CH), 5.02 (d, 1H, J=5.0 Hz, OH), 7.11 Hz (d, J=8.48 Hz, 1H, ArH), 7.35(dd, 2H, J=4.4 Hz, J=1.5 Hz, ArH), 8.55 (dd, J=1.5 Hz, J=4.6 Hz, 2H,ArH).

¹³C NMR (CDCl₃, δ): 24.0, 25.1, 25.8, 25.9, 32.5, 35.8, 41.7, 46.0,46.9, 55.6, 72.6, 121.3 (2×C), 149.2, 149.5 (2×C), 168.9, 173.7.

Preparation of(±)-erythro-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)butan-1-onedihydrochloride Compound 239(±)-cis-(5-Methyl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)-pyrrolidin-1-yl-methanoneEBE 06180

To a stirred solution of KOH (223 mg, 39.7 mmol) in MeOH was added4-acetylpyridine (478 mg, 39.7 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (500 mg, 3.2 mmol). Thereaction mixture was stirred for 3 h at 0° C. and then concentrated. Theresidue was partitioned between EtOAc and H₂O. The organic layer waswashed with brine, dried over MgSO₄, filtered and evaporated to give ayellow oil that was purified by column chromatography (SiO₂) with 20%[v/v] MeOH in EtOAc to give cis- andtrans-(±)-(5-methyl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)-pyrrolidin-1-yl-methanone.The mixture was further purified by column chromatography (SiO₂) using agradient of 2% to 5% [v/v] MeOH [v/v] in CH₂Cl₂ to obtain the purecis-(±)-(5-methyl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)-pyrrolidin-1-yl-methanoneEBE 06180 (122 mg, 51% yield) as white solid.

MW: 259.3; Yield: 51%; White Solid; Mp (° C.): 140.9

R_(f): 0.30 (EtOAc:MeOH=80:20).

¹H-NMR (CDCl₃, δ): 1.45-1.75 (m, 4H, 2×CH₂), 1.81 (s, 3H, CH₃),2.75-2.90 (m, 1H, N—CH₂), 3.10-3.22 (m, 1H, N—CH₂), 3.30-3.40 (t, 2H,J=6.7 Hz, N—CH₂), 4.83 (d, 1H, J=1.7 Hz, NCH), 7.22 (d, 1H, J=1.4 Hz,N═CH), 7.27 (d, 2H, J=6.0 Hz, Ar), 8.57 (d, 2H, J=6.1 Hz, ArH).

¹³C-NMR (CDCl₃, δ): 23.6, 26.0, 27.9, 46.0, 46.5, 77.8, 87.2, 120.2,148.9, 149.6 (2×C), 155.6 (2×C), 165.4.

(±)-erythro-2-Amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)butan-1-onedihydrochloride Compound 239

To a solutioncis-(±)-(5-methyl-5-pyridin-4-yl-4,5-dihydro-oxazol-4-yl)-pyrrolidin-1-yl-methanoneEBE 06180 (50 mg, 0.19 mmol) in MeOH (1 mL) was added a solution of 1 NHCl (1 mL) and the reaction mixture was heated at 60° C. for 2 h. Allthe volatiles were evaporated to give(±)-erythro-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)butan-1-onedihydrochloride Compound 239 (54 mg, 87% yield) as a white solid.

MW: 322.23; Yield: 87%; White Solid; Mp (° C.): 140.9

R_(f): 0.1 (EtOAc:MeOH=80:20, free base).

¹H-NMR (CDCl₃, δ): 1.85-2.05 (m, 7H, CH₃+2×CH₂), 3.35-3.65 (m, 4H,2×N—CH₂), 4.61 (s, 1H, O—CH), 8.23 (d, 2H, J=4.5 Hz, ArH), 8.89 (d, 2H,J=4.3 Hz, ArH).

Preparation of(S)-2-amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 240 tert-Butyl(S)-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamate EBE06190

To a solution of N-Boc-(2S)-3-(pyridin-4-yl)alanine (500 mg, 1.9 mmol)in THF (12 mL) was added N-methylmorpholine (200 μL, 1.9 mmol) and thesolution was stirred for 5 min, cooled at −15° C. and treated dropwisewith isobutyl chloroformate (249 μL, 1.9 mmol). The mixture was stirredfor 10 min and pyrrolidine (1.08 g, 15.2 mmol) was added and allowed towarm to 25° C. with stirring for 3 h. The solvent were removed underreduced pressure and the residue was partitioned between EtOAc andNaH₂PO₄ pH=7.2. The aqueous layer was discarded and the organic layerwas washed with aqueous saturated NaHCO₃, dried over Na₂SO₄, filteredand evaporated. The resulting solid was purified by columnchromatography (SiO₂) with a gradient of 0% to 10% [v/v] MeOH in EtOActo give tert-butyl(S)-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamate EBE06190 (167 mg, 28% yield) as a white solid.

MW: 319.4; Yield: 28%; White Solid; Mp (° C.): 130.0

R_(f): 0.30 (EtOAc:MeOH=90:10).

¹H-NMR (CDCl₃, δ): 1.40 (s, 9H, C(CH₃)₃, 1.65-1.90 (m, 4H, CH₂),2.80-2.95 (m, 1H, CH), 2.95-3.05 (m, 2H, CH₂), 3.30-3.45 (m, 1H, NCH),3.45-3.55 (m, 2H, N—CH₂), 4.60-4.75 (m, 1H, N—CH), 5.42 (d, 1H, J=8.8Hz, NH), 7.16 (dd, 2H, J=4.5 Hz, J=1.5 Hz, ArH), 8.51 (dd, 2H, J=1.5 Hz,J=4.5 Hz, ArH).

¹³C-NMR (CDCl₃, δ): 24.1, 25.8, 28.3 (3×C), 39.1, 45.8, 46.5, 52.6,79.9, 124.8 (2×C), 145.7, 149.8 (2×C), 155.0, 169.2.

(S)-2-Amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 240

To a solution of tert-butyl(S)-1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-2-ylcarbamate EBE06190 (100 mg, 0.313 mmol) in CH₂Cl₂ (3 mL) at 4° C. was added TFA (479μL, 6.26 mmol) and MeOH (0.3 mL) and the reaction was stirred for 2 h.All the volatiles were evaporated to give a product that was treatedwith a suspension of amberlite-400 (OH⁻ form, 5 g) in MeOH. Thesuspension was filtered and washed with MeOH (5×5 mL). The combinedmethanol fractions were evaporated under reduced pressure and thedesired product was isolated by column chromatography (SiO₂) with agradient of 0% to 30% [v/v] MeOH in EtOAc to give(S)-2-amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-one. Theproduct was dissolved in MeOH, cooled at 5° C. and a solution of HCl(0.1 N) (9 mL) was added dropwise. All the volatiles were evaporated togive (S)-2-amino-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 240 (91 mg, 99% yield) as a white solid.

MW: 292.21; Yield: 99%; White Solid; Mp (° C.): 195.9

R_(f): 0.10 (EtOAc:MeOH=90:10, free base).

¹H-NMR (CD₃OD, δ): 1.80-2.05 (m, 4H, CH₂), 3.30-3.40 (m, 5H,N—CH₂+N—CH), 3.60-3.75 (m, 1H, CH), 4.72 (t, 1H, J=7.3 Hz, CH), 8.08 (d,2H, J=5.4 Hz, ArH), 8.87 (d, 2H, J=5.3 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.0, 26.9, 37.2, 47.6, 48.0, 57.2, 130.0 (2×C),143.1 (2×C), 157.3, 166.6.

MS-ESI m/z (% rel. Int.): 220.1 ([MH]⁺, 10), 203.1 (50).

Preparation of(±)-threo-2-amino-3-(6-(trifluoromethyl)pyridin-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 241(±)-trans-(5-(6-(Trifluoromethyl)pyridin-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneEBE 06196

To a solution of KOH (184 mg, 3.28 mmol) in MeOH (10 mL) at 4° C. wasadded 6-(trifluoromethyl)pyridine-3-carbaldehyde (575 mg, 3.28 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (500 mg, 3.28 mmol).The mixture was stirred for 2 h at 4° C. and allowed to warm to 25° C.All the volatiles were evaporated and the resulting product waspartitioned between brine and EtOAc. The organic layer was separated,dried over Na₂SO₄, filtered to give after evaporation of the solvent(±)-trans-(5-(6-(trifluoromethyl)pyridin-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneEBE 06196 (801 mg, 78% yield) as a pale brown solid.

MW: 310.13; Yield: 78%; Pale brown solid.

¹H-NMR (CDCl₃, δ): 1.75-2.10 (m, 4H, 2×CH₂), 3.40-3.60 (m, 3H, 2×N—CH₂),3.90-4.00 (m, 1H, N—CH), 4.57 (dd, 1H, J=8.0 Hz, J=2.2 Hz, N—CH), 6.31(d, 1H, J=8.0 Hz, O—CH), 7.06 (d, 1H, J=2.2 Hz, N═CH), 7.71 (d, 1H,J=8.1 Hz, ArH), 7.84 (dd, 1H, J=1.8 Hz, J=8.1 Hz, ArH), 8.70 (d, 1H,J=1.5 Hz, ArH).

(±)-threo-2-Amino-3-(6-(trifluoromethyl)pyridin-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 241

To a solution of(±)-trans-(5-(6-(trifluoromethyl)pyridin-3-yl)-4,5-dihydrooxazol-4-yl)(pyrrolidin-1-yl)methanoneEBE 06196 (400 mg, 1.28 mmol) in MeOH (2 mL) was added HCl (37%) (10 mL)and the mixture was heated at 60° C. for 2 h with continuous stirring.After evaporation the resulting white solid was treated with asuspension of amberlite-400 (OH⁻ form) in MeOH. The suspension wasfiltered and washed with MeOH (5×5 mL). The combined methanol fractionwere evaporated under reduced pressure and the desired product wasisolated by column chromatography (SiO₂) with a gradient of 0% to 8%[v/v] MeOH in EtOAc to obtain(±)-threo-2-amino-3-(6-(trifluoromethyl)pyridin-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-one(287 mg). To a solution(±)-threo-2-amino-3-(6-(trifluoromethyl)pyridin-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-one(209 mg, 0.69 mmol) in MeOH (2 mL) at 4° C. and treated HCl 37% (10 mL).All the volatiles were evaporated to give(±)-threo-2-amino-3-(6-(trifluoromethyl)pyridin-3-yl)-3-hydroxy-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 241 as a white solid (269 mg, 74% yield).

MW: 376.2; Yield: 74%; White Solid; Mp (° C.): 190.0.

R_(f): 0.3 (EtOAc:MeOH=92:8, free base).

¹H-NMR (CD₃OD, δ): 1.40-1.70 (m, 2H, CH₂), 1.70-1.90 (m, 2H, CH₂),2.35-2.50 (m, 1H, N—CH), 3.15-3.35 (m, 1H, N—CH₂), 3.35-3.45 (m, 2H,N—CH₂), 4.32 (d, 1H, J=8.3 Hz, N—CH), 5.11 (d, 1H, J=8.2 Hz, O—CH), 7.88(d, 1H, J=8.3 Hz, ArH), 8.16 (d, 1H, J=8.0 Hz, ArH), 8.73 (s, 1H, ArH).

¹³C-NMR (CD₃OD, δ): 24.8, 26.6, 47.4, 47.9, 58.7, 71.3, 121.7, 122.9 (d,1C, J=273.2 Hz, CF₃), 137.9, 140.3, 149.0, 149.7, 165.8.

Preparation of(±)-threo-2-amino-3-hydroxy-1-(thiazolidin-3-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 242(±)-threo-2-Amino-3-hydroxy-3-(pyridin-4-yl)propanoic aciddihydrochloride EBE 10038B

To a stirred solution of KOH (2.57 g, 35.4 mmol) in MeOH (35 mL) at 0°C. was added 4-pyridinecarboxaldehyde (3.80 g, 35.4 mmol) andtert-butylisocyano acetate (5 g, 35.4 mmol). The solution was stirredfor 3 h at 0° C. and concentrated to obtain intermediatetrans-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxylate as a pale yellowsolid EBE 10038A.

¹H NMR (CD₃OD, δ): 4.29 (dd, 1H, J=1.7 Hz, J=7.6 Hz, N—CH), 5.67 (d, 1H,J=7.6 Hz, O—CH), 7.29 (d, 1H, J=1.9 Hz, N═CH), 7.46 (d, 2H, J=4.7 Hz,ArH), 8.55 (dd, 2H, J=1.7 Hz, J=4.4 Hz, ArH).

The solid EBE 10038A was dissolved in MeOH (100 mL), stirred 5 min at 0°C. and treated with HCl (12 N) (10.5 mL). The reaction was heated at 60°C. for 2 h, cooled down to 4° C. to form a precipitate that wasfiltered. The filtrate was evaporated and dried to obtain(±)-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)propanoic aciddihydrochloride EBE 10038B with 20% of (±)-erythro isomer (9 g, 99%crude yield) as a pale beige solid.

MW: 255.10; Yield: 99%; Pale Beige Solid.

¹H-NMR (CD₃OD, δ): 3.94 (d, 1H, J=3.9 Hz, N—CH), 4.61 (t, 1H, J=3.0 Hz,O—CH), 8.31 (d, 2H, ArH), 8.95 (m, 2H, ArH). Only the formula and the¹H-NMR description of major threo isomer is shown.

MS-ESI m/z (% rel. Int.): 183.1 ([MH]⁺, 5).

(±)-threo-N-Boc-2-amino-3-(pyridin-4-yl)-3-hydroxy-propionic acid EBE10040

A solution of di-tert-butyldicarbonate (9.28 g, 42.5 mmol) in dioxane(40 mL) was added to a pre-mixed ice cold solution of(±)-threo-2-amino-3-hydroxy-3-(pyridin-4-yl)propanoic aciddihydrochloride EBE 10038B (9.01 g, 35.4 mmol) in a solution of 1 N NaOH(145 mL). The biphasic mixture was stirred at 5° C. for 30 min andallowed to warm to room temperature for 3.5 h, concentrated, cooled inan ice bath, acidified to pH 4-5 and extracted with n-butanol. Thecombined extracts were dried over Na₂SO₄ and filtered to give(±)-threo-N-Boc-2-amino-3-(pyridin-4-yl)-3-hydroxy-propionic acid EBE10040 (3.34 g, 33% yield) as a pale yellow solid.

MW: 282.2; Yield: 33%; Pale Yellow Solid.

¹H-NMR (DMSO-d6, δ): 1.24 (s, 9H, C(CH₃)₃), 4.32 (d, 1H, J=9.4 Hz,N—CH), 5.13 (s, 1H, O—CH), 6.35 (d, 1H, J=9.4 Hz, NH), 7.37 (d, 2H,J=3.8 Hz, ArH), 8.48 (d, 2H, J=3.6 Hz), 20H not seen.

¹³C-NMR (DMSO-d6, δ): 27.9 (3×C), 59.0, 71.3, 78.2, 121.4, 122.8, 148.9,150.3, 151.0, 155.2, 171.6.

MS-ESI m/z (% rel. Int.): 283.2 ([MH]⁺, 10).

HPLC: Method A, detection at 254 nm, EBE 10040 RT=3.17 min, peak area95.9%.

tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(thiazolidin-3-yl)propan-2-ylcarbamateEBE 10042

To a solution of(±)-threo-N-Boc-2-amino-3-(pyridin-4-yl)-3-hydroxy-propionic acid EBE10040 (500 mg, 1.77 mmol) in CH₂Cl₂ (10 mL) at 0° C. was addedtriethylamine (253 μL, 3.54 mmol), hydroxybenzotriazole (239 mg, 1.77mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbo-diimide hydrochloride(EDCI) (340 mg, 1.77 mmol) and thiazolidine (140 μL, 1.77 mmol). Thereaction mixture was stirred for 2 h at 0° C., allowed to warm to roomtemperature, stirred for 16 h and diluted in CH₂Cl₂ (90 mL). The mixturewas washed with brine (3×10 mL), 1 N NaOH (3×10 mL), dried over Na₂SO₄,filtered to give a crude oil that was purified using columnchromatography (SiO₂) with a gradient of 3% to 4% MeOH in CH₂Cl₂. Afterevaporation tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(thiazolidin-3-yl)propan-2-ylcarbamateEBE 10042 (200 mg, 32% yield) was obtained as a yellow oil.

MW: 353.4; Yield: 32%; Yellow Oil.

R_(f): 0.2 (CH₂Cl₂:MeOH=97:3).

¹H-NMR (CDCl₃, δ): 1.29 (s, 9H, (CH₃)₃), 2.90-3.05 (m, 2H, S—CH₂),3.60-3.95 (m, 2H, N—CH₂), 4.40-4.72 (m, 3H, N—CH+S—CH₂—N), 5.10-5.15 (m,1H, OCH), 5.68 (m, 1H, NH), 7.34 (d, 2H, J=6.0 Hz, ArH), 8.53 (d, 2H,J=6.0 Hz, ArH), OH not seen.

¹³C-NMR (CDCl₃, δ), minor rotamer in parenthesis: 28.1 (29.2) [3×C],31.1, (48.5) 48.7, 49.0 (49.1), (56.4) 56.7, 72.3, 80.5, 121.4 [2×C],148.7, 149.4 [2×C], 155.5, (169.0) 169.2.

MS-ESI m/z (% rel. Int.): 354.2 ([MH]⁺, 20).

HPLC: Method A, detection at 254 nm, EBE 10042 RT=3.87 min, peak area98.1%.

(±)-threo-2-Amino-3-hydroxy-1-(thiazolidin-3-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 242

To a solution of tert-butyl(±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(thiazolidin-3-yl)propan-2-ylcarbamateEBE 10042 (150 mg, 424 mg) in MeOH (10 mL) at 4° C. was added a solutionof 1 N HCl in MeOH (12 mL). The reaction mixture was allowed to warm atroom temperature and stirred for 1 h. All the volatiles were evaporatedto give an oily residue that was dissolved in MeOH and treated withEtOAc to form a precipitate. The volatiles were evaporated to give(±)-threo-2-amino-3-hydroxy-1-(thiazolidin-3-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 242 (135 mg, 98% yield) as a pale yellow solid.

MW: 326.24; Yield: 98%; Pale Yellow solid; Mp (° C.): 210.6

¹H-NMR (CDCl₃, δ): 2.90-3.15 (m, 2H, S—CH₂), 3.55-3.90 (m, 1H, N—CH),4.00-4.15 (m, 1H, N—CH), 4.18-4.53 (m, 1H, N—CH), 4.62-4.78 (m, 2H,N—CH₂—S), 5.38-5.49 (m, 1H, O—CH), 8.25 (d, 2H, J=5.7 Hz, 2H, ArH), 8.94(d, 2H, J=5.7 Hz, ArH).

¹³C-NMR (CDCl₃, δ), minor rotamer in parenthesis: (29.9) 31.8, 34.8,(51.1) 52.1, (57.8) 58.2, 71.0 (71.3), 126.8 [2×C], 143.2 [2×C], 161.4,(164.8) 165.2.

MS-ESI m/z (% rel. Int.): 254.2 ([MH]⁺, 15).

Preparation of(±)-threo-2-amino-3-hydroxy-1-(indolin-1-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 243

To a solution of potassiumtrans-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxylate EBE 10038A (500mg, 2.60 mmol) in CH₂Cl₂ (13 mL) were added HOBT (352 mg, 2.60 mmol),EDCI (499 mg, 2.60 mmol) and indoline (292 mL, 2.60 mmol). The reactionmixture was stirred 2 h at 0° C., allowed to warm to RT and stirred for16 h. The reaction mixture was diluted in CH₂Cl₂ (100 mL), wash withbrine (3×25 mL), NaOH 1N (3×25 mL), dried over MgSO₄, filtered to giveafter evaporationtrans-(4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)(1H-indol-1-yl)methanoneSLA 09182 (533 mg, 70% yield) as a pale brown oil. To a solution of SLA09182 (533 mg, 2.6 mmol) in MeOH (12 mL) was added a solution of HCl 37%(880 μL). The reaction was stirred for 3 h at 50° C. and concentratedunder reduced pressure. The resulting product was dissolved in MeOH andtreated with amberlite (OH⁻ form), filtered to give after evaporation aresidue that was purified using silica gel chromatography with agradient of MeOH 0%-10% in CH₂Cl₂ to yield(±)-threo-2-amino-3-hydroxy-1-(indolin-1-yl)-3-(pyridin-4-yl)propan-1-one.The hydrochloride salt was formed by treatment with a solution of 1M HClin MeOH (3.2 mL) to give after evaporation(±)-threo-2-amino-3-hydroxy-1-(indolin-1-yl)-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 243 (149 mg, 16% yield) as a white solid.

MW: 356.25; Yield: 16%; White Solid; Mp (° C.): 202.5

R_(f): 0.30 (CH₂Cl₂: MeOH=90:10, free base)

¹H-NMR (CD₃OD, δ): 3.65-3.80 (m, 1H, N—CH₂), 4.15-4.28 (m, 1H, CH₂),3.65-3.80 (m, 1H, CH₂), 4.15-4.30 (m, 1H, N—CH₂), 4.64 (d, 1H, J=5.4 Hz,N—CH), 5.45 (d, 1H, J=5.3 Hz, O—CH), 6.70 (t, 1H, J=7.4 Hz, ArH), 7.12(dd, 2H, J=7.7 Hz, ArH), 8.07 (d, 1H, J=7.9 Hz, ArH), 8.15 (d, 2H, J=6.2Hz, ArH), 8.79 (d, 2H, J=6.2 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 28.9, 30.8, 58.4, 71.1, 118.5, 126.1, 126.4, 126.5(2×C), 128.4, 133.6, 142.9, 143.6 (2×C), 161.0, 164.7.

MS-ESI m/z (% rel. Int.): 284.2 ([MH]⁺, 10).

Preparation of(±)-threo-2-amino-3-(3,5-dichloropyridin-4-yl)-3-hydroxy-1-(2H-pyrrol-1(5H)-yl)propan-1-onedihydrochloride Compound 245trans-(5-(3,5-Dichloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 09022

To a stirred and cooled (0° C.) solution of KOH (0.315 g, 5.62 mmol) inmethanol (70 mL) was added a mixture of3,5-dichloropyridine-4-carbaldehyde (0.989 mg, 5.62 mmol) and2-isocyano-1-(2H-pyrrol-1(5H)-yl)ethanone SLA 07178 (0.696 g, 5.11mmol). The solution was stirred 2 h with continued cooling and thenconcentrated. The residue was partitioned between EtOAc (50 mL) andwater (50 mL). The organic layer was combined with additional EtOAcextracts (3×50 mL), washed with brine (70 mL) and dried with MgSO₄,filtered and evaporated to obtain a crude product which was purified bycolumn chromatography (florisil, EtOAc:cyclohexane=80:20) to obtainafter evaporationtrans-(5-(3,5-dichloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 09022 (1.267 g, 80% yield) as a pale yellow solid.

MW: 312.15; Yield: 80%; Pale Yellow Solid.

R_(f): 0.15 (EtOAc: cyclohexane=80:20).

¹H-NMR (CDCl₃, δ): 4.30-4.32 (m, 3H, 1.5×CH₂), 4.77-4.83 (m, 1H,0.5×CH₂), 4.86 (dd, 1H, J=2.2 Hz, J=8.8 Hz, CH—N), 5.84-5.88 (m, 2H,CH═CH), 6.86 (d, 1H, J=8.7 Hz, CH—O), 6.96 (d, 1H, J=2.2 Hz, O—CH═N),8.52 (s, 2H, ArH).

¹³C-NMR (CDCl₃, δ): 52.1, 52.6, 59.2, 75.1, 123.9, 124.4, 131.5, 139.5,147.6, 148.3, 151.0, 153.9, 164.3.

(±)-threo-2-Amino-3-(3,5-dichloropyridin-4-yl)-3-hydroxy-1-(2H-pyrrol-1(5H)-yl)propan-1-onedihydrochloride Compound 245

To a stirred solution oftrans-(5-(3,5-dichloropyridin-4-yl)-4,5-dihydrooxazol-4-yl)(2H-pyrrol-1(5H)-yl)methanoneSLA 09022 (1.26 g, 4.04 mmol) in methanol (20 mL) was added HCl 37% (1.5mL). The reaction mixture was stirred for 3 h at RT, concentrated andthe resulting yellow oil was co evaporated twice with EtOAc andtriturated with EtOAc to obtain after filtration and drying under vacuum(±)-threo-2-amino-3-(3,5-dichloropyridin-4-yl)-3-hydroxy-1-(2H-pyrrol-1(5H)-yl)propan-1-onedihydrochloride (1.13 g, 75% yield) as a pale yellow solid.

MW: 375.16; Yield: 75%; Pale yellow solid; Mp (° C.): 176.5

R_(f): 0.15 (CH₂Cl₂: MeOH=90:10, free base).

¹H-NMR (CD₃OD, δ): 3.30-3.32 (m, 1H, 0.5×CH₂), 3.90-3.97 (m, 1H,0.5×CH₂), 4.25-4.37 (m, 2H, CH₂), 4.76 (d, 1H, J=10.3 Hz, N—CH),5.60-5.65 (m, 1H, ═CH), 5.70 (d, 1H, J=10.3 Hz, O—CH) 5.78-5.82 (m, 1H,═CH), 8.57 (s, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 56.6, 56.7, 57.3, 72.4, 128.0, 128.9, 136.3 (2C),145.8, 152.5 (2C), 168.1.

Preparation ofN-(1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)prop-2-en-2-yl)formamidehydrochloride (cis:trans isomers mixture) Compound 246

To a stirred solution of triphenylphosphine (200 mg, 0.76 mmol) in 5 mLof CH₃CN were added at 20° C. diethylazodicarbonate (120 μl, 0.76 mmol),Et₃N (55 μl, 0.38 mmol) andN-((±)-threo-1-hydroxy-3-oxo-1-(pyridin-4-yl)-3-(pyrrolidin-1-yl)propan-2-yl)formamidehydrochloride Compound 216 (115 mg, 0.38 mmol). The mixture was stirred2 h at 70° C. then solvent was evaporated. The obtained residue waspurified by column chromatography (SiO₂, EtOAc:MeOH=9:1) to giveN-(1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)prop-2-en-2-yl)formamideTTA 08074A (80 mg, 43% yield). HCl treatment in EtOAc with HCl 0.4 N indiethyl ether (1 mL, 0.4 mmol) gave after evaporation and dryingN-(1-oxo-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)prop-2-en-2-yl)formamidehydrochloride (trans:cis isomers mixture) Compound 246 (60 mg, 28%yield) as a pale yellow pasty product.

MW: 281.74; Yield: 28%; Pale Yellow Pasty Product.

R_(f): 0.24 (EtOAc:MeOH=9:1).

¹H-NMR (CD₃OD, δ): 1.80-2.05 (bs, 4H, 2×CH₂), 3.25-3.45 (bs, 2H, CH₂—N),3.55-3.70 (bs, 2H, CH₂—N), 6.87 (s, 1H, CH), 7.82 (d, 2H, J=5.0 Hz,ArH), 8.31 (s, 1H, HC═O), 8.65 (d, 2H, J=5.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.1, 26.5, 47.1, 48.9, 109.0, 125.6 (2×C), 127.7,142.2 (2×C), 142.6, 154.9, 161.6.

MS-ESI m/z (% rel. Int.): 246.1 ([MH]⁺, 5), 175.1 (100).

HPLC: Method A, detection UV 254 nm, Compound 246 RT=1.90 min, peak area95.0%.

Preparation of (2S,3R)- &(2R,3S)-2-amino-1-((R)-3-hydroxypyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compound 2471-((R)-3-hydroxypyrrolidin-1-yl)-2-isocyanoethanone VIB 01172

To stirred and cooled (0° C.) methyl isocyanoacetate (96% technicalgrade, 1.7 g, 17.21 mmol) was slowly added (R)-(+)-3-pyrrolidinol (1.5g, 17.21 mmol) and MeOH (5 mL). The mixture was stirred for 3 h at RTand concentrated. Brine was added (30 mL) and the mixture was extractedwith EtOAc (3×50 mL), dried over MgSO₄, filtered and evaporated toobtained crude 1-((R)-3-hydroxypyrrolidin-1-yl)-2-isocyanoethanone VIB01172 (1.3 g, 49% yield) as a yellow solid.

MW: 154.17; Yield: 49%; Yellow Solid; Mp (° C.)=55.9

¹H-NMR (CDCl₃, δ): 1.95-2.20 (m, 2H, CH₂), 2.60-2.82 (m, 1H, OH),3.30-3.68 (m, 4H, 2×CH₂), 4.20-4.36 (m, 2H, CH₂), 4.49-4.65 (m, 1H,CH—O).

MS-ESI m/z (% rel. Int.): 155.1 ([MH]⁺, 90).

trans-((R)-3-hydroxypyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01174

To a stirred and cooled (0° C.) solution of KOH (0.40 g, 7.13 mmol) inMeOH (8 mL) were added successively pyridine-4-carbaldehyde (0.84 g,7.84 mmol) and 1-((R)-3-hydroxypyrrolidin-1-yl)-2-isocyanoethanone VIB01172 (1.10 g, 7.13 mmol). The mixture was stirred at 0° C. to RT for 24h. After evaporation of MeOH, the mixture was partitioned between EtOAc(50 mL) and H₂O (10 mL). The aqueous layer was further extracted withEtOAc (2×50 mL). The EtOAc fractions were combined, washed twice withbrine (2×10 mL), dried over MgSO₄, filtered and evaporated. Afterevaporation and drying trans-((R)-3-hydroxypyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01174(490 mg, diastereoisomeric mixture in ratio 1:1, 26% yield) wereobtained as a crude pale yellow solid.

MW: 261.28; Yield: 26%; Pale Yellow Solid.

¹H NMR (CDCl₃, δ): 1.88-2.22 (m, 2H, CH₂), 3.50-3.80 (m, 3H, 1.5×CH₂),3.95-4.20 (m, 1H, 0.5×CH₂), 4.40-4.65 (m, 2H, CH—N & CH—O), 4.74 (s, 1H,OH), 6.18-6.22 (m, 1H, CH—O), 7.00-7.12 (m, 1H, HC═N), 7.20-7.30 (m, 2H,ArH), 8.52-8.68 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 262.2 ([MH]⁺, 45), 235.2 (75), 148 (100).

(2S,3R)- &(2R,3S)-2-Amino-1-((R)-3-hydroxypyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compound 247

To a solution of trans-((R)-3-hydroxypyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones VIB 01174(0.49 g, 1.87 mmol) in methanol (6.5 mL) was added hydrochloric acid 37%(575 μL). After heating (50° C.) the mixture for 2 h the reactionmixture was concentrated and the crude product was coevaporated twicewith EtOAc. After trituration with EtOAc, filtration and drying (2S,3R)-&(2R,3S)-2-amino-1-((R)-3-hydroxypyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides (420 mg, diastereoisomeric mixture in ratio 1:1, 69%yield) were obtained as a pale pink solid.

MW: 324.2; Yield: 69%; Pale Pink Solid; Mp (° C.): 177.0

¹H-NMR (CD₃OD, δ): 1.88-2.22 (m, 2H, CH₂), 2.70-3.80 (m, 4H, 2×CH₂),4.20-4.65 (m, 2H, CH—O & CH—N), 5.20-5.45 (m, 1H, CH—O), 8.10-8.25 (m,2H, ArH), 8.80-9.00 (m, 2H, ArH), 2×OH & NH₂ not seen.

MS-ESI m/z (% rel. Int.): 252.2 ([MH]⁺, 37), 235.1 (63), 148.0 (100).

Preparation of(−)-threo-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 248 and(+)-threo-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 249 Extraction of the Free Base:

(2S,3R)- &(2R,3S)-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochlorides Compound 214 (300 mg, 0.92 mmol) were dissolved in 10mL of a Na₂CO₃ (10%) solution and the aqueous mixture was then saturatedwith NaCl. The aqueous phase was extracted by 5×15 mL of a mixtureCH₂Cl₂:2-PrOH (9:1). The organic phase was dried over MgSO₄ andevaporated to afford 163 mg (70%) of the corresponding free base ofCompound 214.

Analytical Chiral Separation:

20 μL of a 1 mg/mL solution of Compound 214 were injected on ChiralpakAD: flow-rate=1 mL/min, temperature=25° C., mobile phasehexane:ethanol=7:3, detection on UW 220 nm and on polarimeter, firsteluted diastereoisomer Compound 248 Rt1(−)=20.94 min, second eluteddiastereoisomer Rt2(+)=24.77 min, k1(−)=5.93, k2(+)=7.20, α=1.21 andresolution Rs=1.21. The integration of the UV signal gives 42% for thefirst diastereoisomer compound 248 and 58% for the second Compound 249(the UV response is different for the two diastereoisomers).

Semi-Preparative Chiral Separation:

170 mg of the free base of Compound 214 were dissolved in 6 mL ofethanol, and 30 μL of this solution were injected every 9 min onChiralpak AD-H, flow-rate=2 mL/min, mobile phase hexane:ethanol=7:3,detection on UV 220 nm. 195 successive injections were done. The twomain fractions were identified on UV and collected in two differentflasks. The solvent was removed in vacuo at 30° C. The resulting solidwas dissolved in 50 mL of CH₂Cl₂ and then filtered on a 0.45 μmmillipore membrane. After evaporation of CH₂Cl₂, the solid was dissolvedin 50 mL of methanol and then filtered. For the free base of the firstdiastereoisomer, a new series of injections was needed to remove twoUV-visible impurities collected in the same flask: in the samechromatographic conditions, 30 injections of 100 μL of a 25 mg/mLsolution were made every 20 min. The salts Compound 248 and Compound 249were regenerated according to the same procedure reported for Compound203 and Compound 204.

The regenerated salts Compound 248 and Compound 249 were injected inanalytical conditions, the diastereoisomeric excesses for Compound 248and Compound 249 were determined to be higher than 96%.

(−)-threo-2-Amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 248

MW: 326.19; 70 mg obtained; Pale Yellow Solid; Mp (° C.): toohygroscopic.

Diastereoisomeric excess >96% measured by HPLC at 220 nm (Chiralpak AD).

α²⁵ _(D)=−2.0 (methanol, c=1).

¹H-NMR (CD₃OD, δ): 1.85-2.38 (m, 2H, CH₂), 2.72-4.05 (m, 4H, 2×CH₂),4.49-4.62 (m, 1H, CH—N), 5.10-5.48 (m, 2H, CH—O & CH—F), 8.11-8.25 (m,2H, ArH), 8.82-8.98 (m, 2H, ArH).

(+)-threo-2-Amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-4-yl)propan-1-onedihydrochloride Compound 249

MW: 326.19; 85 mg obtained; Pale Yellow Solid; Mp (° C.): toohygroscopic.

Diastereoisomeric excess >96% measured by HPLC at 220 nm (Chiralpak AD).

α²⁵ _(D)=+31.7 (methanol, c=1).

¹H-NMR (CD₃OD, δ): 1.82-2.38 (m, 2H, CH₂), 2.90-4.00 (m, 4H, 2×CH₂),4.35-4.60 (m, 1H, CH—N), 5.00-5.48 (m, 2H, CH—O & CH—F), 8.11-8.25 (m,2H, ArH), 8.82-9.00 (m, 2H, ArH).

Preparation of(±)-threo-2-Amino-N-ethyl-3-hydroxy-N-methyl-3-pyridin-4-yl-propanamidedihydrochloride Compound 250trans-N-ethyl-4,5-dihydro-N-methyl-5-(pyridin-4-yl)oxazole-4-carboxamideSLA 09190

To a solution of potassiumtrans-4,5-dihydro-5-(pyridin-4-yl)oxazole-4-carboxylate EBE 10038A (501mg, 2.60 mmol) in CH₂Cl₂ (12 mL) were added HOBT (352 mg, 2.60 mmol),EDCI (500 mg, 2.60 mmol) and N-methylethanamine (223 mL, 2.60 mmol). Thereaction mixture was stirred 2 h at 0° C., allowed to warm to roomtemperature and stirred for 16 h. The reaction mixture was diluted inCH₂Cl₂ (100 mL), washed with brine (2×25 mL), 1 N NaOH (2×25 mL), driedover MgSO₄, filtered to give after evaporationtrans-N-ethyl-4,5-dihydro-N-methyl-5-(pyridin-4-yl)oxazole-4-carboxamideSLA 09190 (144 mg, 24% yield) as a pale brown oil.

MW: 233.27; Yield: 24%; Pale Brown Oil.

¹H NMR (CDCl₃, δ): 1.15-1.30 (m, 3H, CH₃), 3.22 (s, 3H, CH₃—N),3.40-3.80 (m, 2H, N—CH₂), 4.59 (dd, 1H, J=2.2 Hz, J=7.8 Hz, N—CH), 6.24(d, J=7.7 Hz, OCH), 7.02 (d, 1H, J=1.0 Hz, N═CH), 7.23 (d, 2H, J=4.8 Hz,ArH), 8.61 (d, 2H, J=4.6 Hz, ArH).

MS-ESI m/z (% rel. Int.): 234.2 ([MH]⁺, 30).

(±)-threo-2-Amino-N-ethyl-3-hydroxy-N-methyl-3-pyridin-4-yl-propanamidedihydrochloride Compound 250

To a solution ofN-ethyl-4,5-dihydro-N-methyl-5-(pyridin-4-yl)oxazole-4-carboxamide SLA09190 (144 mg, 0.6 mmol) in MeOH (5 mL) was added a solution of HCl 37%(240 μL). The reaction was stirred for 3 h at 50° C. and concentratedunder reduced pressure. The resulting product was dissolved in MeOH andtreated with amberlite (OH⁻ form), filtered to give after evaporation aresidue that was purified by column chromatography (SiO₂, with agradient of MeOH 10% in CH₂Cl₂) to yield to2-amino-N-ethyl-3-hydroxy-N-methyl-3-pyridin-4-yl-propionamide. Thehydrochloride salt was formed by treatment of this free base with asolution of HCl 1 M in MeOH (1 mL) to give after evaporation2-amino-N-ethyl-3-hydroxy-N-methyl-3-pyridin-4-yl-propanamidedihydrochloride Compound 250 as a pale yellow solid (80 mg, 44% yield).

MW: 296.19; Yield: 44%; Pale Yellow Solid; Mp (° C.): 114.7

R_(f): 0.30 (CH₂Cl₂:MeOH=90:10, freebase).

¹H-NMR (CD₃OD, δ): 0.96-1.10 (m, 3H, CH₃), 2.81 (s, 1.8H major rotamer,0.6×CH₃), 2.88 (s, 1.2H minor rotamer, 0.4CH₃), 3.18-3.55 (m, 2H, CH₂),4.66 (d, 0.4H minor rotamer, J=6.6 Hz, 0.4×N—CH), 4.69 (d, 0.6H majorrotamer, J=6.3 Hz, 0.6×N—CH), 5.24 (d, 0.4H minor rotamer, J=6.9 Hz,0.4×O—CH), 5.27 (d, 0.6H major rotamer, J=6.3 Hz, 0.6×O—CH), 8.08 (t,2H, J=6.5 Hz, ArH), 8.86 (d, 2H, J=5.0 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 12.0, (13.6), (33.4), 35.3, 44.5, (45.6), 56.1,(56.2), 71.6, (72.1), 126.2 (2×C), 144.5, 144.6, 159.6, 166.5. ( ) Minorrotamer in parenthesis.

Preparation of(2R,3S)-2-(3,4-dichlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 251

To a solution of(−)-(2R,3S)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 203 (175 mg, 0.57 mmol) and Et₃N (175 μL, 1.25mmol) in MeOH (5 mL) in a 50 mL round bottom flask equipped with amagnetic stirrer and under nitrogen atmosphere was added slowly at RT3,4-dichlorobenzaldehyde (112 mg, 0.63 mmol). The reaction mixture wasstirred at RT for 20 h. Then AcOH (65 μL, 1.15 mmol) and NaBH₃CN (50 mg,0.74 mmol) were added. The reaction mixture was stirred at RT foranother 15 h. MeOH was evaporated and EtOAc (100 mL) was added. Theorganic phase was washed with a mixture of saturated sodium carbonate (5mL) and brine (20 mL), then with brine (10 mL) and dried over MgSO₄,filtered and evaporated. The crude product was purified by columnchromatography (SiO₂, eluent EtOAc:MeOH=95:5) to give an oil(−)-(2R,3S)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-one(182 mg, 81% yield). This free base (182 mg, 0.46 mmol) was dissolved inMeOH (2 mL) at 4° C. and a solution of HCl 0.1 N in isopropanol (10.2mL, 1.01 mmol) was added. After evaporation at 30° C., a mixture ofEtOAc:MeOH=95:5 was added to yield, after evaporation and drying, to(2R,3S)-2-(3,4-dichlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 251 as a white solid (208 mg, 78% yield).

MW: 467.22; Yield: 78%; White Solid; Mp (° C.): 195.1

R_(f): 0.22 (EtOAc:MeOH=95:5, free base).

¹H-NMR (CD₃OD, δ): 1.55-1.77 (m, 4H, 2×CH₂), 2.46-2.53 (m, 1H,0.5×N—CH₂), 3.20-3.30 (m, 3H, 1.5×N—CH₂), 4.25 (d, 1H, J=13.3 Hz,0.5×N—CH₂), 4.38 (d, 1H, J=13.3 Hz, 0.5×N—CH₂), 4.52 (d, 1H, J=7.7 Hz,N—CH), 5.33 (d, 1H, J=7.7 Hz, O—CH), 7.48 (dd, 1H, J=8.3 Hz, J=1.7 Hz,ArH), 7.62 (dd, 1H, J=8.3 Hz, J=1.2 Hz, ArH), 7.74 (s, 1H, ArH), 8.15(d, 2H, J=5.6 Hz, ArH), 8.91 (d, 2H, J=5.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 24.7, 26.6, 47.6, 48.2, 50.4, 64.0, 72.2, 126.8(2×C), 131.7, 132.2, 132.3, 133.8, 133.9, 135.1, 143.5 (2×C), 160.6,163.8.

MS-ESI m/z (% rel. Int.): 394.1/396.1/398.1 ([MH]⁺, 60/45/10), 219.2(100).

HPLC: Method A, detection UV 254 nm, Compound 251 RT=3.83 min, peak area99.5%.

Preparation of(2S,3R)-2-(3,4-dichlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 252

Same experimental as for Compound 251 preparation starting from(+)-(2S,3R)-2-amino-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 204 (175 mg, 0.57 mmol). After purification bycolumn chromatography(2S,3R)-2-(3,4-dichlorobenzylamino)-3-hydroxy-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)propan-1-onewas obtained (187 mg, 83% yield). This free base (187 mg, 0.47 mmol) wasdissolved in MeOH (2 mL) at 4° C. and a solution of HCl 0.1 N inisopropanol (10.4 mL, 1.04 mmol) was added. After evaporation at 30° C.,a mixture of EtOAc:MeOH=95:5 was added to yield, after evaporation anddrying, to Compound 252 as a white solid (212 mg, 80% yield).

MW: 467.22; Yield: 80%; White Solid; Mp (° C.): 187.5

R_(f): 0.22 (EtOAc:MeOH=95:5, free base).

¹H-NMR (CDCl₃, δ): 1.55-1.77 (m, 4H, 2×CH₂), 2.46-2.53 (m, 1H,0.5×N—CH₂), 3.20-3.30 (m, 3H, 1.5×N—CH₂), 4.25 (dd, 1H, J=13.3 Hz,0.5×N—CH₂), 4.40 (d, 1H, J=13.3 Hz, 0.5×N—CH₂), 4.52 (d, 1H, J=7.7 Hz,N—CH), 5.33 (d, 1H, J=7.4 Hz, O—CH), 7.48 (d, 1H, J=8.3 Hz, ArH), 7.62(dd, 1H, J=8.3 Hz, J=0.8 Hz, ArH), 7.74 (s, 1H, ArH), 8.15 (d, 2H, J=5.7Hz, ArH) 8.91 (d, 2H, J=5.7 Hz, ArH).

¹³C-NMR (CD₃OD, *): 24.7, 26.6, 47.6, 48.2, 50.4, 64.0, 72.3, 126.8(2×C), 131.7, 132.2, 132.3, 133.8, 133.9, 135.1, 143.5 (2×C), 160.6,163.8.

MS-ESI m/z (% rel. Int.): 394.1/396.1/398.1 ([MH]⁺, 60/45/10), 219.2(100).

HPLC: Method A, detection UV 254 nm, Compound 252 RT=3.83 min, peak area99.5%.

Preparation of (E)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)prop-2-en-1-onehydrochloride Compound 253

To a solution of 3-(4-pyridinyl)acrylic acid (1.01 g, 6.77 mmol) inCHCl₃ (20 mL) in a 100 mL round bottom flask equipped with a magneticstirrer and under nitrogen atmosphere was added 1-hydroxybenzotriazole(1.11 g, 8.21 mmol). The reaction mixture was stirred at RT for 10 min.Then 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (1.56 g, 8.15 mmol)was added. The reaction mixture was stirred at 4° C. for 10 min. Thenpyrrolidine (1.11 mL, 18.3 mmol) was added slowly and the reactionmixture was stirred for 15 h at +4° C. to RT. Dichloromethane (200 mL)was added and organic phase was washed with brine (100 mL), a solutionof NaOH 0.5 N (100 mL) and brine (50 mL). The organic phase was driedover MgSO₄, filtered, and evaporated to obtain(E)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)prop-2-en-1-one (1.30 g, 94%yield). This free base (1.3 g, 6.40 mmol) was dissolved in MeOH (10 mL)at 4° C. and a solution of HCl 0.1 N in isopropanol (79 mL, 7.9 mmol)was added. After evaporation at 30° C. and drying,(E)-3-(pyridin-4-yl)-1-(pyrrolidin-1-yl)prop-2-en-1-one hydrochlorideCompound 253 was obtained as a beige solid (1.40 g, 87% yield).

MW: 238.71; Yield: 87%; Beige Solid; Mp (° C.): 229.4

R_(f): 0.35 (EtOAc:MeOH=95:5, free base).

¹H-NMR (CD₃OD, δ): 1.92-2.10 (m, 4H, 2×CH₂), 3.56 (t, 2H, J=6.7 Hz,N—CH₂), 3.80 (t, 2H, J=6.7 Hz, N—CH₂), 7.59 (d, 1H, J=15.6 Hz, CH═C),7.68 (d, 1H, J=15.6 Hz, CH═C), 8.35 (d, 2H, J=5.7 Hz, ArH) 8.86 (d, 2H,J=5.6 Hz, ArH).

¹³C-NMR (CD₃OD, δ): 25.2, 27.0, 47.6, 48.2, 126.7 (2×C), 131.9, 136.5,143.1 (2×C), 154.5, 164.6.

MS-ESI m/z (% rel. Int.): 203.2 ([MH]⁺, 100).

HPLC: Method A, detection UV 254 nm, Compound 253 RT=3.18 min, peak area99.5%.

Preparation of(±)-threo-2-amino-3-hydroxy-3-(1-methyl-1H-imidazol-2-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 254trans-(4,5-Dihydro-5-(1-methyl-1H-imidazol-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneLPO 01190B

To a stirred and cooled (0° C.) solution of potassium hydroxide (0.33 g,5.0 mmol) in MeOH (6 mL) were added a mixture of 1-methyl-2-imidazolecarboxaldehyde (0.56 g, 5.0 mmol) and2-isocyano-1-(pyrrolidin-1-yl)ethanone BLE 04098 (0.70 g, 5.0 mmol). Thesolution was stirred 3 h at 4° C. and then concentrated. The residue waspartitioned between EtOAc (100 mL) and water (20 mL). The organic layerwas washed with brine (10 mL) and dried over MgSO₄, filtered andevaporated. Concentration afforded a crude product which was purified bycolumn chromatography (florisil, EtOAc:MeOH=95:5 to 90:10) to yield,after evaporation and drying, totrans-(4,5-dihydro-5-(1-methyl-1H-imidazol-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneLPO 01190B (0.32 g, 25% yield) as a brown oil.

MW: 248.28; Yield: 25%; Brown Oil.

R_(f): 0.30 (EtOAc:MeOH=9:1, free base).

¹H-NMR (CDCl₃, δ): 1.94-2.12 (m, 4H, 2×CH₂), 3.50 (t, 2H, J=6.5 Hz,N—CH₂), 3.69-4.13 (m, 5H, N—CH₂, N—CH₃), 5.68 (dd, 1H, J=7.7 Hz, J=2.3Hz, CH—N), 6.19 (d, 1H, J=7.7 Hz, CH—O), 6.82 (d, 1H, J=2.2 Hz, CH═N),6.94 (d, 1H, J=1.1 Hz, ArH), 7.00 (d, 1H, J=1.1 Hz, ArH).

¹³C-NMR (CDCl₃, δ): 24.2, 26.0, 32.9, 46.4, 46.7, 71.2, 72.7, 123.0,127.9, 143.7, 153.7, 166.6.

MS-ESI m/z (% rel. Int.): 267.3 ([MH+18]⁺, 10), 196.2 (100).

HPLC: Method A, detection UV 254 nm, LPO 01190B RT=3.92 min, peak area99.5%.

(±)-threo-2-Amino-3-hydroxy-3-(1-methyl-1H-imidazol-2-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 254

A solution oftrans-(4,5-dihydro-5-(1-methyl-1H-imidazol-2-yl)oxazol-4-yl)(pyrrolidin-1-yl)methanoneLPO 01190B (320 mg, 1.29 mmol) and HCl 37% (0.4 mL, 13 mmol) in MeOH (6mL) was stirred at 50° C. for 3 h in a 50 mL round bottom flask. Thesolvent was evaporated and the product was precipitated by a mixture ofMeOH:EtOAc:Et₂O=3:12:5 (20 mL). Solvents were evaporated at 30° C. togive, after evaporation and drying,(±)-threo-2-amino-3-hydroxy-3-(1-methyl-1H-imidazol-2-yl)-1-(pyrrolidin-1-yl)propan-1-onedihydrochloride Compound 254 as a pale yellow solid (305 mg, 76% yield).

MW: 311.21; Yield: 76%; Pale Yellow Solid; Mp (° C.): 183.4

R_(f): 0.30 (CH₂Cl₂:MeOH=95:5, free base).

¹H-NMR (CD₃OD, δ): 1.75-1.99 (m, 4H, 2×CH₂), 2.80-2.88 (m, 1H,0.5×N—CH₂), 3.30-3.70 (m, 3H, 1.5×N—CH₂), 3.95 (s, 3H, N—CH₃), 4.62 (d,1H, J=8.3 Hz, N—CH), 5.51 (d, 1H, J=8.3 Hz, O—CH), 7.66 (s, 2H, ArH).

¹³C-NMR (CD₃OD, δ): 24.9, 26.9, 36.0, 47.9 (2×C), 56.2, 65.1, 121.1,126.5, 145.1, 164.4.

MS-ESI m/z (% rel. Int.): 239.3 ([MH]⁺, 10), 134.1 (100).

HPLC: Method A, detection UV 254 nm, RT=0.8 min, peak area 99.5%.

Preparation of (2S,3R)- &(2R,3S)-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-3-yl)propan-1-onedihydrochlorides Compound 255trans-((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones SLA 11014

To a stirred and cooled 0° C. solution of KOH (0.216 mg, 4.22 mmol) inmethanol (5 mL) was added1-((S)-3-fluoropyrrolidin-1-yl)-2-isocyanoethanone VIB 01166 (0.600 g,4.22 mmol) and pyridine-3-carbaldehyde (0.40 mL, 3.84 mmol). Thesolution was stirred for 20 h at 0° C. After evaporation under reducedpressure, the residue obtained was partitioned between EtOAc and H₂O.The product was extracted with EtOAc (4×50 mL) and washed with brine (25mL), dried over MgSO₄, filtered and evaporated to yield to a productthat was purified using chromatography (florisil, EtOAc:MeOH=95:5),trans-((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones SLA 11014were obtained as a yellow solid (0.464 g, diastereoisomeric mixture inratio about 1:1, 46% yield).

MW: 263.27; Yield: 46%; Yellow Solid; Mp (° C.)=171.7

R_(f): 0.25 (EtOAc:MeOH=95:5).

¹H-NMR (CDCl₃, δ): 1.85-2.45 (m, 2H, CH₂), 3.50-4.10 (m, 3H, CH₂ &N—CH), 4.25-4.65 (m, 2H, N—CH₂), 5.15-5.25 (m, 0.5H, 0.5×CHF), 5.35-5.45(m, 0.5H, 0.5×CHF), 6.21 (d, 1H, J=7.6 Hz, O—CH), 7.04 (d, 1H, J=2.1 Hz,N═CH), 7.30-7.38 (m, 1H, ArH), 7.60-7.68 (m, 1H, ArH), 8.55-8.65 (m, 2H,ArH).

MS-ESI m/z (% rel. Int.): 264.1 ([MH]⁺, 18).

(2S,3R)- &(2R,3S)-2-Amino-1-((5)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-3-yl)propan-1-onedihydrochlorides Compound 255

To a solution of trans-((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(pyridin-4-yl)oxazol-4-yl)methanones SLA 11014(0.450 g, 1.71 mmol) in methanol (40 mL) was added HCl 37% (5 mL). Afterheating at 50° C. for 3 h, the mixture was concentrated and the crudeproduct was co-evaporated twice with EtOAc. Trituration with EtOAc andfiltration yielded, after drying, to (2S,3R)- &(2R,3S)-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(pyridin-3-yl)propan-1-onedihydrochlorides Compound 255 (540 mg, diastereoisomeric mixture inratio about 1:1, 97% yield) as a yellow solid.

MW: 326.19; Yield: 97%; Yellow Solid; Mp (° C.): 168.9

¹H-NMR (CD₃OD, δ): 1.85-2.40 (m, 2H, CH₂), 3.45-4.20 (m, 4H, 2×CH₂),4.40-4.75 (m, 1H, N—CH), 5.30-5.60 (m, 2H, O—CH & CHF), 8.15-8.25 (m,1H, ArH), 8.70-8.80 (m, 1H, ArH), 8.90-9.10 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 254.1 ([MH]⁺, 81.38), 236.2 (25).

((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)methanones SLA 11016

To a stirred and cooled 0° C. solution of KOH (0.216 mg, 3.85 mmol) inmethanol (8 mL) was added1-((S)-3-fluoropyrrolidin-1-yl)-2-isocyanoethanone VIB 01166 (0.600 g,4.22 mmol) and thiophene-3-carbaldehyde (0.37 mL, 3.85 mmol). Thesolution was stirred for 20 h at 0° C. After evaporation under reducedpressure, the residue obtained was partitioned between EtOAc and H₂O.The product was extracted with EtOAc (4×50 mL) and washed brine (25 mL),dried over MgSO₄, filtered and evaporated to yield to a product that waspurified using chromatography (florisil, gradient EtOAc:MeOH=95:5 to85:15). ((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- and(4R,5S)-4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)methanones SLA 11016were obtained as a yellow solid (0.411 g, diastereoisomeric mixture inratio about 1:1, 32% yield).

MW: 268.31; Yield: 32%; Yellow Solid; Mp (° C.)=132.9

R_(f): 0.35 (EtOAc:MeOH=80:20).

¹H-NMR (CDCl₃, δ): 1.80-2.45 (m, 2H, CH₂), 3.50-4.10 (m, 3H, CH—N &CH₂), 4.20-4.70 (m, 2H, CH₂), 5.15-5.45 (m, 1H, CHF), 6.18-6.25 (m, 1H,O—CH), 6.99 (d, 1H, J=2.2 Hz, N═CH), 7.00-7.15 (m, 1H, ArH), 7.28-7.35(m, 1H, ArH), 7.32-7.40 (m, 1H, ArH).

MS-ESI m/z (% rel. Int.): 269.0 ([MH]⁺, 10).

(2S,3R)- &(2R,3S)-2-Amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(thiophen-3-yl)propan-1-onedihydrochlorides Compound 256

To a solution of ((S)-3-fluoropyrrolidin-1-yl)((4S,5R)- &(4R,5S)-4,5-dihydro-5-(thiophen-3-yl)oxazol-4-yl)methanones SLA 11016(0.400 g, 1.49 mmol) in methanol (50 mL) was added hydrochloric acid 37%(4 mL). After heating at 50° C. for 3 h, the mixture was concentratedand the crude product was co-evaporated twice with EtOAc. Triturationwith EtOAc and filtration and drying afforded (2S,3R)- &(2R,3S)-2-amino-1-((S)-3-fluoropyrrolidin-1-yl)-3-hydroxy-3-(thiophen-3-yl)propan-1-onedihydrochlorides Compound 256 (451 mg, diastereoisomeric mixture about1:1, 91% yield) as a yellow solid.

MW: 331.23; Yield: 91%; Yellow Solid; Mp (° C.): 221.6

¹H-NMR (CD₃OD, δ): 1.25-2.05 (m, 2H, CH₂), 2.10-2.50 (m, 1H, 0.5×CH₂),3.20-3.65 (m, 3H, 1.5×CH₂), 3.90-4.10 (m, 1H, CH—N), 4.70-5.10 (m, 2H,O—CH & CHF), 6.92-6.99 (m, 1H, ArH), 7.21-7.32 (m, 2H, ArH).

MS-ESI m/z (% rel. Int.): 259.1 ([MH]⁺, 25).

1. A compound of the formula

where R₁ is H or alkyl of 1 to 6 carbons, R₂ is H, alkyl of 1 to 6carbons or the R₁ and R₂ groups together with the nitrogen form asaturated or unsaturated 4, 5, 6 or 7 membered ring that optionallyincludes one or two heteroatoms independently selected from N, O and S,said 4, 5, 6 or 7 membered ring optionally being substituted with one ortwo COOH, CH₂OH, OH, B(OH)₂, halogen groups, cyano groups, or with oneor two alkyl groups having 1 to 6 carbons, or one or two carbons of saidrings being attached to an oxygen to form keto groups and said 4, 5, 6or 7 membered ring optionally being condensed with an aromatic ornon-aromatic 5 or 6 membered ring that optionally includes 1 orheteroatoms selected from N, O and S; R₃ is independently selected fromH, alkyl of 1 to 20 carbons, cycloalkyl of 3 to 6 carbons, aryl orheteroaryl, aryl-alkyl where the alkyl moiety has 1 to 4 carbons,aryl-(hydroxy)alkyl where the alkyl moiety has 1 to 4 carbons,heteroaryl-alkyl where the alkyl moiety has 1 to 4 carbons, orhetero-(hydroxy)alkyl where the alkyl moiety has 1 to 4 carbons, saidaryl or heteroaryl groups being optionally substituted with 1 to 3groups independently selected from the group consisting of halogen,alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3carbons, or R₃ is CO—R₇, SO₂R₇ or CO—O—R₇ where R₇ is H, alkyl of 1 to 1to 20 carbons, alkyl of 1 to 20 carbons substituted with and NH₂ groupor with an NH—COalkyl group where the alkyl group has one to 6 carbons,aryl or heteroaryl, aryl-alkyl where the alkyl moiety has 1 to 4 carbonsor heteroaryl-alkyl where the alkyl moiety has 1 to 4 carbons, said arylor heteroaryl groups being optionally substituted with 1 to 3 groupsindependently selected from the group consisting of halogen, alkyl of 1to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3 carbons; R₄is H, alkyl of 1 to 6 carbons or CO—R₈ where R₈ is alkyl of 1 to 6carbons; the wavy lines represent bonds connected to carbons having R orS configuration; the dashed lines represent a bond or absence of a bondwith the proviso that the ring containing the dashed lines is aromatic;m, n and q are integers independently selected from 0, 1, 2 or 3 withthe proviso that the sum of m, n and q is 2 or 3; s is zero (0) or whenX is N then s is zero (0) or 1; W, X and Y independently represent a CH,CR₅, CR₆ or a heteroatom selected independently of N, O and S, and R₅and R₆ are independently selected from H, halogen, alkyl of 1 to 6carbons, halogen substituted alkyl of 1 to 6 carbons, alkoxy of 1 to 6carbons and thioxy of 1 to 3 carbons, phenyl, or R₅ and R₆ together withthe atoms to which they are attached jointly form a carbocyclic or aheterocyclic ring, the carbocyclic ring having 5 or 6 atoms in the ring,the heterocyclic ring having 5 or 6 atoms in the ring and 1 to 3heteroatoms independently selected from N, O and S; said carbocyclic orheterocyclic ring jointly formed by R₅ and R₆ being optionallysubstituted with 1 to 6 R₉ groups where R₉ is independently selectedfrom halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons andthioxy of 1 to 3 carbons or a pharmaceutically acceptable salt of saidcompound with the proviso that Formula 1 does not cover compounds whereR₄ is H, R₁ and R₂ jointly with the nitrogen form a pyrrolidino ormorpholino ring, the sum of m, n and q is 3, and where none of W, X andY represent a heteroatom with the further proviso that the formula doesnot cover the compounds of the formula below


2. A compound in accordance with claim 1 where the sum of the integersm, n and q is
 3. 3. A compound in accordance with claim 2 where one ofthe W, X and Y groups is N.
 4. A compound in accordance with claim 3where R₅ and R₆ together with the atoms to which they are attachedjointly form a carbocyclic or a heterocyclic ring.
 5. A compound inaccordance with claim 2 where none of the W, X and Y groups is aheteroatom.
 6. A compound in accordance with claim 5 where R₅ and &together with the atoms to which they are attached jointly form acarbocyclic or a heterocyclic ring.
 7. A compound in accordance withclaim 1 where the sum of the integers m, n and q is 2 and at least oneof W, X and Y is a heteroatom.
 8. A compound in accordance with claim 7where only one of W, X and Y represents a heteroatom.
 9. A compound inaccordance with claim 7 where two of the W, X and Y groups eachrepresent an independently selected heteroatom.
 10. A compound inaccordance with claim 1 where R₄ is H.
 11. A compound in accordance withclaim 10 where both R₃ groups are H.
 12. A compound in accordance withclaim 10 where one R₃ group is H and the other R₃ group is CO—R₇ oralkyl of 1 to 20 carbons.
 13. A compound in accordance with claim 10where R₃ is independently selected from H and alkyl of 1 to 10 carbons.14. A compound in accordance with claim 1 where R₄ is CO—R₈.
 15. Acompound in accordance with claim 1 where R₁ and R₂ together with thenitrogen form a 4, 5, 6 or 7 membered ring.
 16. A compound in accordancewith claim 15 where R₁ and R₂ together with the nitrogen form a 5membered ring.
 17. A compound in accordance with claim 15 where R₁ andR₂ together with the nitrogen form a 6 membered ring.
 18. A compound inaccordance with claim 1 where both R₅ and R₆ are hydrogen.
 19. Acompound of the formula

where R₁ is H or alkyl of 1 to 6 carbons, R₂ is H, alkyl of 1 to 6carbons or the R₁ and R₂ groups together with the nitrogen form asaturated or unsaturated 4, 5, 6 or 7 membered ring that optionallyincludes one or two heteroatoms independently selected from N, O and S,said 4, 5, 6 or 7 membered ring optionally being substituted with one ortwo COOH, CH₂OH, OH, B(OH)₂, halogen groups, cyano groups or with one ortwo alkyl groups having 1 to 6 carbons; R₃ is H, CO—R₇ or CO—O—R₇ whereR₇ is H, alkyl of 1 to 1 to 20 carbons, cycloalkyl of 3 to 6 carbons,aryl or heteroaryl, aryl-alkyl, aryl(hydroxy)alkyl, heteroaryl-alkyl orheteroalkyl(hydroxy)alkyl where the alkyl moiety has 1 to 4 carbons,said aryl or heteroaryl groups being optionally substituted with 1 to 3groups independently selected from the group consisting of halogen,alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3carbons; R₄ is H, alkyl of 1 to 6 carbons, or CO—R₈ where R₈ is alkyl of1 to 6 carbons, R₅ and R₆ are independently selected from H, halogen,alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3carbons, or R₅ and R₆ together with the atoms to which they are attachedjointly form a carbocyclic or a heterocyclic ring, the carbocyclic ringhaving 5 or 6 atoms in the ring, the heterocyclic ring having 5 or 6atoms in the ring and 1 to 3 heteroatoms independently selected from N,O and S; said carbocyclic or heterocyclic ring jointly formed by R₅ andR₆ being optionally substituted with 1 to 6 R₉ groups where R₉ isindependently selected from halogen, alkyl of 1 to 6 carbons, alkoxy of1 to 6 carbons, and the wavy lines represent bonds of the alpha or betaconfiguration, or any pharmaceutically acceptable salt of said compound.20. A compound in accordance with claim 19 where 4 is H.
 21. A compoundin accordance with claim 19 where both R₃ groups are H.
 22. A compoundin accordance with claim 19 where both R₅ and R₆ are hydrogen.
 23. Acompound in accordance with claim 22 having the formula

or a pharmaceutically acceptable salt of said compound.
 24. A compoundin accordance with claim 23 where R₄ is H.
 25. A compound in accordancewith claim 23 where both R₃ groups are H.
 26. A compound in accordancewith claim 23 that has the formula

or a pharmaceutically acceptable salt of said compound.
 27. A compoundin accordance with claim 23 that has the formula

or a pharmaceutically acceptable salt of said compound.
 28. A compoundin accordance with claim 23 having the formula

or any other pharmaceutically acceptable salt of said compound.
 29. Acompound in accordance with claim 23 having the formula

or any other pharmaceutically acceptable salt of said compound.
 30. Acompound in accordance with claim 23 having the formula

or any other pharmaceutically acceptable salt of said compound.
 31. Acompound in accordance with claim 23 having the formula

or any other pharmaceutically acceptable salt of said compound.
 32. Acompound in accordance with claim 23 having the formula

or any other pharmaceutically acceptable salt of said compound.
 33. Acompound in accordance with claim 23 having the formula

or any other pharmaceutically acceptable salt of said compound.
 34. Acompound in accordance with claim 23 that has the formula

or any other pharmaceutically acceptable salt of said compound.
 35. Acompound in accordance with claim 23 that has the formula

or any other pharmaceutically acceptable salt of said compound.
 36. Acompound in accordance with claim 23 that has the formula

or any other pharmaceutically acceptable salt of said compound.
 37. Acompound in accordance with claim 23 that has the formula

or a pharmaceutically acceptable salt of said compound.
 38. A compoundin accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 39. Acompound in accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 40. Acompound in accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 41. Acompound in accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 42. Acompound in accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 43. Acompound in accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 44. Acompound in accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 45. Acompound in accordance with claim 19 having the formula

or any other pharmaceutically acceptable salt of said compound.
 46. Acompound in accordance with claim 45 having the formula

or any other pharmaceutically acceptable salt of said compound.
 47. Acompound in accordance with claim 45 having the formula

or any other pharmaceutically acceptable salt of said compound.
 48. Acompound of the formula

where R₁ is H or alkyl of 1 to 6 carbons, R₂ is alkyl of 1 to 6 carbonsor the R₁ and R₂ groups together with the nitrogen form a saturated orunsaturated 4, 5, 6 or 7 membered ring that optionally includes one ortwo heteroatoms independently selected from N, O and S, said 4, 5, 6 or7 membered ring optionally being substituted with one or two COOH,CH₂OH, OH, B(OH)₂, halogen groups, cyano groups, or with one or twoalkyl groups having 1 to 6 carbons, R₃ is H, CO—R₇ or CO—O—R₇ where R₇is H, alkyl of 1 to 1 to 20 carbons, aryl or heteroaryl, cycloalkyl of 3to 6 carbons, aryl-alkyl aryl(hydroxy)alkyl, heteroaryl-alkyl orheteroaryl(hydroxy)alkyl where the alkyl moiety has 1 to 4 carbons, saidaryl or heteroaryl groups being optionally substituted with 1 to 3groups independently selected from the group consisting of halogen,alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3carbons; R₄ is H, alkyl of 1 to 6 carbons, or CO—R₈ where R₈ is alkyl of1 to 6 carbons; R₅ and R₆ are independently selected from H, halogen,alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons, thioxy of 1 to 3carbons and phenyl, or R₅ and R₆ together with the atoms to which theyare attached jointly form a carbocyclic or a heterocyclic ring, thecarbocyclic ring having 5 or 6 atoms in the ring, the heterocyclic ringhaving 5 or 6 atoms in the ring and 1 to 3 heteroatoms independentlyselected from N, O and S; said carbocyclic or heterocyclic ring jointlyformed by R₅ and R₆ being optionally substituted with 1 to 6 R₉ groupswhere R₉ is independently selected from halogen, alkyl of 1 to 6carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3 carbons, and thewavy lines represent bonds connected to carbons having R or S ofconfiguration, with the proviso that the claim does not cover compoundswhere R₄ is H, R₁ and R₂ jointly with the nitrogen form a pyrrolidino ormorpholino ring, or any pharmaceutically acceptable salt of saidcompound.
 49. A compound in accordance with claim 48 having the formula

or any other pharmaceutically acceptable salt of said compound.
 50. Acompound of the formula

where X is O or S; R₁ is H or alkyl of 1 to 6 carbons, R₂ is H, alkyl of1 to 6 carbons or the R₁ and R₂ groups together with the nitrogen form asaturated or unsaturated 5 or 6 membered ring that optionally includesone or two heteroatoms independently selected from N, O and S, said 4,5, 6 or 7 membered ring optionally being substituted with one or twoCOOH, CH₂OH, OH, B(OH)₂, halogen groups or cyano groups with one or twoalkyl groups having 1 to 6 carbons, R₃ is H, CO—R₇ or CO—O—R₇ where R₇is H, alkyl of 1 to 1 to 20 carbons, aryl or heteroaryl, aryl-alkyl,aryl(hydroxy)alkyl, heteroaryl-alkyl or heteroaryl(hydroxy)alkyl wherethe alkyl moiety has 1 to 4 carbons, said aryl or heteroaryl groupsbeing optionally substituted with 1 to 3 groups independently selectedfrom the group consisting of halogen, alkyl of 1 to 6 carbons, alkoxy of1 to 6 carbons and thioxy of 1 to 3 carbons; R₄ is H, alkyl of 1 to 6carbons, or CO—R₈ where R₈ is alkyl of 1 to 6 carbons; R₅ and R₆ areindependently selected from H, halogen, alkyl of 1 to 6 carbons, alkoxyof 1 to 6 carbons and thioxy of 1 to 3 carbons, or R₅ and R₆ togetherwith the atoms to which they are attached jointly form a carbocyclic ora heterocyclic ring, the carbocyclic ring having 5 or 6 atoms in thering, the heterocyclic ring having 5 or 6 atoms in the ring and 1 to 3heteroatoms independently selected from N, O and S; said carbocyclic orheterocyclic ring jointly formed by R₅ and R₆ being optionallysubstituted with 1 to 6 R₉ groups where R₉ is independently selectedfrom halogen, alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons, and thewavy lines represent bonds connected to carbons having R or Sconfiguration, or any pharmaceutically acceptable salt of said compoundwith the further proviso that the claim does not cover the compoundshown below


51. A compound in accordance with claim 50 having the formula

or a pharmaceutically acceptable salt of said compound.
 52. A compoundin accordance with claim 51 having the formula

or a pharmaceutically acceptable salt of said compound.
 53. A compoundin accordance with claim 51 having the formula

or any other pharmaceutically acceptable salt of said compound.
 54. Acompound in accordance with claim 53 having the formula

or any other pharmaceutically acceptable salt of said compound.
 55. Acompound in accordance with claim 53 having the formula


56. A method of treating a mammal in need of such treatment with apharmaceutical composition having an analgesic effect, the compositionincluding a compound in accordance with claim
 1. 57. A method inaccordance with claim 56, for treating a mammal in need of suchtreatment with a pharmaceutical composition having an analgesic effect,the composition including a compound in accordance with claim
 34. 58. Amethod in accordance with claim 56, for treating a mammal in need ofsuch treatment with a pharmaceutical composition having an analgesiceffect, the composition including a compound in accordance with claim36.
 59. A method of stimulating the immune system of a mammal in need ofsuch treatment with a pharmaceutical composition having animmunostimulant effect, the composition including a compound inaccordance with claim
 1. 60. A method in accordance with claim 59 forstimulating the immune system of a mammal in need of such treatment witha pharmaceutical composition having an immunostimulant effect, thecomposition including a compound in accordance with claim
 35. 61. Amethod in accordance with claim 59 for stimulating the immune system ofa mammal in need of such treatment with a pharmaceutical compositionhaving an immunostimulant effect, the composition including a compoundin accordance with claim
 37. 62. A compound of the formula

where R₁ is H or alkyl of 1 to 6 carbons, R₂ is H, alkyl of 1 to 6carbons or the R₁ and R₂ groups together with the nitrogen form asaturated or unsaturated 4, 5, 6 or 7 membered ring that optionallyincludes one or two heteroatoms independently selected from N, O and S,said 4, 5, 6 or 7 membered ring optionally being substituted with one ortwo COOH, CH₂OH, OH, B(OH)₂, halogen or cyano groups groups or with oneor two alkyl groups having 1 to 6 carbons, or one or two carbons of saidrings being attached to an oxygen to form keto groups and said 4, 5, 6or 7 membered ring optionally being condensed with an aromatic ornon-aromatic 5 or 6 membered ring that optionally includes 1 orheteroatoms selected from N, O and S; R₃ is independently selected fromH, alkyl of 1 to 20 carbons, cycloalkyl of 3 to 6 carbons, aryl orheteroaryl, aryl-alkyl where the alkyl moiety has 1 to 4 carbons,aryl-(hydroxy)alkyl where the alkyl moiety has 1 to 4 carbons,heteroaryl-alkyl where the alkyl moiety has 1 to 4 carbons orhetero-(hydroxy)alkyl where the alkyl moiety has 1 to 4 carbons, saidaryl or heteroaryl groups being optionally substituted with 1 to 3groups independently selected from the group consisting of halogen,alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3carbons, or R₃ is CO—R₇, SO₂R₇ or CO—O—R₇ where R₇ is H, alkyl of 1 to 1to 20 carbons, alkyl of 1 to 20 carbons substituted with and NH₂ groupor with an NH—COalkyl group where the alkyl group has one to 6 carbonsaryl or heteroaryl, aryl-alkyl where the alkyl moiety has 1 to 4 carbonsor heteroaryl-alkyl where the alkyl moiety has 1 to 4 carbons, said arylor heteroaryl groups being optionally substituted with 1 to 3 groupsindependently selected from the group consisting of halogen, alkyl of 1to 6 carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3 carbons; thewavy lines represent bonds connected to carbons having R or Sconfiguration; the dashed lines represent a bond or absence of a bondwith the proviso that the ring containing the dashed lines is aromatic;R₉ and R₁₀ are one of the combinations selected from the groupconsisting of (1) R₉ is NOR₁₁ and R₁₀ does not exist, (2) R₉ is OR₁₁ andR₁₀ is alkyl of 1 to 6 carbons, (3) when the dashed lines betweencarbons 2 and 3 of the propionic acid moiety represent a bond then R₉ isH or alkyl of 1 to 6 carbons and R₁₀ does not exist; R₁₁ is H, alkyl of1 to 6 carbons or CO—R₁₂ where R₁₂ is alkyl of 1 to 6 carbons; m, n andq are integers independently selected from 0, 1, 2 or 3 with the provisothat the sum of m, n and q is 2 or 3; s is zero (0) or when X is N thens is zero (0) or 1; W, X and Y independently represent a CH, CR₅, CR₆ ora heteroatom selected independently of N, O and S, and R₅ and R₆ areindependently selected from H, halogen, alkyl of 1 to 6 carbons, halogensubstituted alkyl of 1 to 6 carbons, alkoxy of 1 to 6 carbons and thioxyof 1 to 3 carbons, phenyl, or R₅ and R₆ together with the atoms to whichthey are attached jointly form a carbocyclic or a heterocyclic ring, thecarbocyclic ring having 5 or 6 atoms in the ring, the heterocyclic ringhaving 5 or 6 atoms in the ring and 1 to 3 heteroatoms independentlyselected from N, O and S; said carbocyclic or heterocyclic ring jointlyformed by R₅ and R₆ being optionally substituted with 1 to 6 R₁₂ groupswhere R₁₂ is independently selected from halogen, alkyl of 1 to 6carbons, alkoxy of 1 to 6 carbons and thioxy of 1 to 3 carbons or apharmaceutically acceptable salt of said compound.
 63. A compound inaccordance with claim 62 where the sum of the integers m, n and q is 3.64. A compound in accordance with claim 62 where one of the W, X and Ygroups is N.
 65. A compound in accordance with claim 62 where the R₁ andR₂ groups together with the nitrogen form a 5-membered ring.
 66. Acompound in accordance with claim 62 where R₉ is NOR₁₁ and R₁₀ does notexist.
 67. A compound in accordance with claim 62 where R₉ is methyl andR₁₀ is OR₁₁.
 68. A compound in accordance with claim 62 where the dashedline between carbons 2 and 3 represents a bond.